Background/Aim: Chemotherapy resistance in triple-negative breast cancer (TNBC) cells is well doc... more Background/Aim: Chemotherapy resistance in triple-negative breast cancer (TNBC) cells is well documented. Therefore, it is necessary to develop safer and more effective therapeutic agents to enhance the outcomes of chemotherapeutic agents. The natural alkaloid sanguinarine (SANG) has demonstrated therapeutic synergy when coupled with chemotherapeutic agents. SANG can also induce cell cycle arrest and trigger apoptosis in various cancer cells. Materials and Methods: In this study, we investigated the molecular mechanism underlying SANG activity in MDA-MB-231 and MDA-MB-468 cells as two genetically different models of TNBC. We employed various assays including Alamar Blue to measure the effect of SANG on cell viability and proliferation rate, flow cytometry analysis to study the potential of the compound to induce apoptosis and cell cycle arrest, quantitative qRT PCR apoptosis array to measure the expression of different genes mediating apoptosis, and the western system was used to analyze the impact of the compound on AKT protein expression. Results: SANG lowered cell viability and disrupted cell cycle progression in both cell lines. Furthermore, S-phase cell cycle arrest-mediated apoptosis was found to be the primary contributor to cell growth inhibition in MDA-MB-231 cells. SANG-treated TNBC cells showed significantly up-regulated mRNA expression of 18 genes associated with apoptosis, including eight TNF receptor superfamily (TNFRSF), three members of the BCL2 family, and two members of the caspase (CASP) family in MDA-MB-468 cells. In MDA-MB-231 cells, two members of the TNF superfamily and four members of the BCL2 family were affected. The western study data showed the inhibition of AKT protein expression in both cell lines concurrent with up-regulated BCL2L11 gene. Our results point to the AKT/PI3K signaling pathway as one of the key mechanisms behind SANG-induced cell cycle arrest and death. Conclusion: SANG shows anticancer properties and apoptosis-related gene expression changes in the two TNBC cell lines and suggests AKT/PI3K pathway implication in apoptosis induction and cell cycle arrest. Thus, we propose SANG's potential as a solitary or supplementary treatment agent against TNBC. In the United States, breast cancer (BC) is the most common malignancy and the second leading cause of death among women aged 20-59 years (1). Gene expression is controlled by molecular signal transduction, and alterations in transcriptional settings distinguish cancer cells (2). BC is a heterogeneous cancer type categorized based on its molecular features. Triple-negative breast cancer (TNBC) affects approximately 15% of patients with BC (2). However, the incidence of this subtype is two to three folds higher in African American (AA) women than in other ethnic groups (3, 4). TNBC is the most aggressive BC and has a poor outcome compared to different BC subtypes (3, 5). Based on immunohistochemical characteristics, the expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) are missing in TNBC cells (3, 6). Currently, chemotherapy is the principal treatment for patients with TNBC. However, disease recurrence can manifest within the first two years 323
International Journal of Environmental Research and Public Health
Being cognizant of the pronounced health advantages of breastfeeding for both the nursing mother ... more Being cognizant of the pronounced health advantages of breastfeeding for both the nursing mother and her infant, the breastfeeding dyad, we examined breastfeeding rates among Floridian women who gave birth from 2012 to 2014 (N = 639,052). We investigated the associations between breastfeeding initiation and WIC-based breastfeeding support (the Special Supplemental Nutrition Program for Women, Infants, and Children), education level, and race and ethnicity. We compared the percentage of breastfeeding mothers between those in the WIC program to those who were not, and we compared breastfeeding rates across racial and ethnic groups. Consistent with previous reports, black newborns in this study were breastfed at lower rates than other racial groups, and WIC program participants were less likely to breastfeed than non-WIC program participants. However, by breaking down the data by education level and race, and ethnicity, we see a significantly increased rate of breastfeeding due to WIC ...
Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit carcinogenesis in... more Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit carcinogenesis in cancer cells. We have previously shown DATS's ability to decrease the percentage of viable cells, inhibit cell migration and modulate genes involved in the nuclear factor kappa-light-chainenhancer of activated B cells (NF-ĸB) and mitogen-activated protein kinase (MAPK) signaling. Materials and Methods: This study aimed to compare the efficacy of DATS in tumor necrosis factor alpha (TNF-α) induced MDA-MB-231 and MDA-MB-468 cells and investigate its role in cell-death signaling via cell cycle, flow cytometry, and caspase assay. Results: DATS exhibit a time-dependent accumulation of G 2 /M phase cells in both cell lines, with higher effects in the MDA-MB-468 for all time points. DATS's ability to decrease the percentage of viable cells in both MDA-MB-231 and MDA-MB-468 cells was shown by a significant but slight increase of early and late apoptosis in the presence of DATS compared to control. Moreover, MDA-MB-468 cells showed more sensitivity to the DATS effect, evidenced by the higher percentage of apoptosis than MDA-MB-231 cells. The caspase studies showed a significant increase in caspase 3 and 8 activity in the presence of DATS, compared to control, in both cell lines. DATS showed no significant increase in caspase 9 activity in both cell lines compared to the control. Conclusion: DATS-induced apoptosis in human breast cancer cells is mediated, at least in part, by cell cycle arrest and caspase activity. These findings provide information for future studies into the role of DATS in TNBC therapy and prevention.
ASPET 2023 Annual Meeting Abstract - Drug Discovery and Development
ID 14491 Poster Board 296 For thousands of years, humans have been orally consuming herbal botani... more ID 14491 Poster Board 296 For thousands of years, humans have been orally consuming herbal botanical medicines, ascribing the beneficial effects to the plant itself. Oddly, we have not yet realized the impact of ingesting dead/live microorganisms in these plants. The residual dead remains of the herbal microbiome can and do leave a trail of thermal and pH-stable immune-stimulating antigenic epitopes with the capacity to act in the human gut. The current study employs a high throughput screening to elucidate the immune-stimulating properties of over-the-counter (OTC) / point-of-sale (POS) herbs. The findings illustrate the existence of an immune active herbal bioactive microbiome, which is plant specific but independent of the plant itself. In this study, an initial high throughput screening of 2535 phytochemicals and OTC/POS'S botanicals was carried out to elucidate pro-inflammatory effects in RAW 264.7 macrophages, with many as potent as positive controls: lipopolysaccharide (LPS) (E.coli 0111:B4). Microbial pathogen-associated molecular pattern molecules (PAMPS) in bio-active herbs were removed from the plants by spin column application, extracts were retested for a bioactive response, and the pH stability of LPS analyzed using a stomach simulation model. Lastly, we reverse cultured the "residual remains" in 7 OTC/POS herbs which were isolated, colonized, and sequenced using Illumina targeting the V4 region of the 16S rRNA gene and the V1-V3 region of the 18S rRNA gene. The obtained results show that Out of 2535 compounds screened, not a single phytochemical or drug (>500 tested) could mimic LPS, which was only observed for 65 "specific" herbal medicine plants, having high concentrations of PAMPS, dominating in roots and oceanic marine sea kelps /algae's [e.g.
ASPET 2023 Annual Meeting Abstract - Cancer Pharmacology
ID 54503 Poster Board 446 Triple-negative breast cancer (TNBC) has historically had fewer treatme... more ID 54503 Poster Board 446 Triple-negative breast cancer (TNBC) has historically had fewer treatment alternatives than other breast cancer types. Therefore, it is critical to explore and pinpoint potential biomarkers that could be utilized in treating TNBC. By doing this, we will significantly enhance a patient's prognosis and quality of life. Apoptosis-controlling genes could be manipulated to increase the death of cancer cells. It is essential to comprehend how the genes involved in the apoptotic pathway interact to identify potential therapeutic targets. Therefore, the current study is aimed to evaluate the gene expression, protein-protein interaction (PPI), and transcription factor interaction of 27 apoptosis-regulated genes in TNBC using integrated bioinformatics methods to assess the gene expression, protein-protein interaction (PPI) and transcription factor interaction. Our findings demonstrated that CASP2, CASP3, DAPK1, TNF, TRAF2, and TRAF3 were substantially associated with the overall survival rate (OS) difference of TNBC patients. The results also show that breast tissue gene expressions of BNIP3, TNFRSF10B, MCL1, and CASP4 were downregulated compared to normal breast tissue in UALCAN. At the same time, BIK, AKT1, BAD, FADD, DIABLO, and CASP9 were downregulated in bc-GeneEx-Miner v4.5 mRNA expression (BCGM) databases. Based on the GO term enrichment analysis, the Cellular process (GO:0009987), which has about 21 apoptosis-regulated genes, is the top category in the biological process (BP), followed by biological regulation (GO:0065007). We identified 29 pathways, including p53, angiogenesis, and apoptosis signaling. We examined the PPIs between the genes that regulate apoptosis; CASP3 and CASP9 interact with FADD, MCL1, TNF, TNFRSRF10A, and TNFRSF10; additionally, CASP3 significantly forms PPIs with CASP9, DFFA, and TP53, while CASP9 with DIABLO. In the top 10 transcription factors, the androgen receptor (AR) interacts with five apoptosis-regulated genes (p<0.0001; q<0.01), followed by RARA (Retinoic Acid Receptor Alpha) (p<0.0001; q<0.01) and RNF2 (Ring Finger Protein) (p<0.0001; q<0.01). Overall, the gene expression profile, the PPIs, and the apoptosis-TF interaction findings suggested that the 27 apoptosis-regulated genes might be used as promising targets in treating and managing TNBC. Furthermore, from a total of 27 key genes, CASP2, CASP3, DAPK1, TNF, TRAF2, and TRAF3 were significantly correlated with poor overall survival in TNBC (p-value < 0.05), which could play essential roles in the progression of TNBC and provide attractive therapeutic targets that may offer new candidate molecules for targeted therapy. It was concluded from this study that genetic alterations altering apoptosis could be possible biomarkers for the prognosis and facilitate the search for alternative treatment targets for TNBC.
Muscadine grapes accumulate higher amounts of bioactive phenolics compared with other grape speci... more Muscadine grapes accumulate higher amounts of bioactive phenolics compared with other grape species. To identify the molecular events associated with polyphenolic accumulation that influence antioxidant capacity, two contrasting muscadine genotypes (C5 and C6) with varied phenolic/flavonoid content and antioxidant activity were investigatedviaRNA-sequencing during berry development. The results showed that berry development is concomitant with transcriptome profile changes, which was more pronounced at the véraison (V) stage. Despite that the downregulation pattern of gene expression dominated the upregulation through berry development, the C5 genotype maintained higher expression levels. Comparative transcript profiling allowed the identification of 94 differentially expressed genes with potential relevance in regulating fruit secondary metabolism, including 18 transcription factors and 76 structural genes. The genes underlying the critical enzymes in the modification reactions of ...
Muscadine berries display enhanced nutraceutical value due to the accumulation of distinctive phy... more Muscadine berries display enhanced nutraceutical value due to the accumulation of distinctive phytochemical constituents with great potential antioxidant activity. Such nutritional and health merits are not only restricted to muscadine, but muscadine berries accumulate higher amounts of bioactive polyphenolics compared with other grape species. For the genetic study of the antioxidant trait in muscadine, a multi-locus genome-wide association study (GWAS) with 350 muscadine genotypes and 1,283 RNase H2 enzyme-dependent amplicon sequencing (rhAmpSeq) markers was performed. Phenotyping was conducted with several antioxidant-related traits, including total phenolic content (TPC), total flavonoid content (TFC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, and FRAP antioxidant assay in muscadine berry skin. The correlation coefficient analysis revealed that the TPC, and DPPH/FRAP activities were significantly correlated. Through the GWAS analysis, 12 QTNs were id...
A distinctive tumor microenvironment associated with chronic inflammation, cancer development, an... more A distinctive tumor microenvironment associated with chronic inflammation, cancer development, and drug resistance in triple-negative breast cancer (TNBC) affect disproportionally Caucasian and African American women. PURPOSE: This investigation presents possible new biomarkers that could be modulated by the natural compound pentagalloyl glucose (PGG) in MM-231 (Caucasians) and MM-468 (African American) TNBC cells. METHODS: Cytokine arrays, ELISA, RT-PCR, flow cytometry, and Western analysis were performed. RESULTS: PGG reduced the expression of GRO-α/CXCL1 pro-inflammatory cytokine, which is associated with tumor recurrence and chemoresistance. PGG inhibited IκBKE and MAPK1 gene and protein expression indicating a possible signaling pathway for CXCL1 downregulation through NFκB and MAPK signaling. PGG also inhibited cell growth in both cell lines, which could be associated with a decrease in CXCL1 levels. Besides, PGG induced apoptosis, modulating numerous genes, including caspases...
Rosmarinic acid (RA) is a polyphenolic compound with various pharmacological properties, includin... more Rosmarinic acid (RA) is a polyphenolic compound with various pharmacological properties, including anti-inflammatory, immunomodulatory, anti-mutagenic, antioxidant, and anticancer activities. This study evaluated the effects of RA in two racially different triple-negative breast cancer (TNBC) cell lines. One cell line was derived from Caucasian origin (MDA-MB-231), and another one was derived from an African American origin (MDA-MB-468). The apoptotic effect of RA was performed using An Annexin V-FITC Apoptosis Detection Kit, while the Propidium Iodide Flow Cytometry Kit was used to detect the cell cycle distribution. Gene expression profiling was established using Human Apoptosis PCR array. The results obtained show that RA significantly caused cytotoxic and antiproliferative effects in both cell lines in a dose- and time-dependent manner. It was noted that RA induced cell cycle arrest-related apoptosis and altered the expression of many apoptosis-involved genes differently. In MDA...
Background/Aim: Clear-cell renal cell carcinoma (ccRCC) is the most common and aggressive form of... more Background/Aim: Clear-cell renal cell carcinoma (ccRCC) is the most common and aggressive form of all urological cancers, with poor prognosis and high mortality. Despite growing evidence of involvement in carcinogenesis, the role of KRAB-ZFP in ccRCC has not been fully explored. KRAB Zinc finger proteins (KRAB-ZFPs) are the largest family of mammalian transcription regulators. They are differentially expressed in various tissues during cellular development and phenotypic differentiation. Materials and Methods: In this study, the levels of transcripts of ccRCC from The Cancer Genome Atlas (TCGA) dataset were used to identify prognostic biomarkers in this disease. Results: Using bioinformatics techniques, we demonstrate that approximately 60% of KRAB zinc finger proteins located on chromosome 19p13.2 are differentially expressed, with all but two being down-regulated in ccRCC. Moreover, ZNF844, a paralog of ZNF433, was the most down-regulated across all histological grades and pathological stages (p<0.001). In addition, the decrease in ZNF844 expression was associated with poor patient survival (HR=0.41; 95% CI=0.3-0.56; p<0.0001). Gene Set Enrichment Analysis of genes inversely co-expressed with ZNF844 revealed that enriched pathways were consistently related to immune and translation processes (p<0.05, FDR <0.05). Lastly, ZNF844 expression showed moderate, inverse correlation to Helper Tcell (CD4 or Th1) subtype 1 (R=-0.558, p=5.15×10-39) infiltration and with the exhausted T-cell phenotype (R=-0.37; p=4.1×10-21). Conclusion: Down-regulation of KRAB-ZFPs at 19p13.2 may represent a signature for ccRCC. Moreover, ZNF844 is a prognostic marker for ccRCC and may serve as a putative immune-related tumor suppressor gene. Clear cell renal carcinoma (ccRCC) is the most common neoplastic disease affecting the kidneys. It accounts for at least 70% of all kidney cancers (1, 2). In advanced stages, ccRCC becomes resistant to conventional therapies (1-3). Moreover, later stages are associated with poor prognosis with a 5-year overall survival rate of less than 10% (1-3). Despite growing evidence of involvement in carcinogenesis, the role of KRAB-ZFP in ccRCC has not been fully explored. Kruppel-associated box domain-containing zinc-finger proteins (KRAB-ZFP) represent the most prominent family of transcriptional repressors (4, 5). The human genome contains over 300 KRAB-ZFPs genes that produce more than 800 transcripts that encode proteins, pseudogenes, and splice variants (6-8). KRAB-ZFPs are differentially expressed in various tissues during cellular development. Several of these genes are involved in embryogenesis, genome imprinting, and biological processes that determine phenotypes in stem cells (9-11). Given their roles in cellular differentiation and their ability to alter gene expression via epigenetic processes and chromatin remodeling, it is conceivable that dysregulation of KRAB-ZFP transcription factors could facilitate tumorigenesis and tumor progression. Interestingly, several large KRAB-ZFP clusters are located on chromosome 19 (chr19), and several studies have linked genetic alterations in chromosome 19p13 to cancers (12-18). Using data mining techniques, we recently identified ZNF433, a zinc finger that is differentially expressed in ccRCC (19) and that may be a biomarker for ccRCC. ZNF433 is part of a cluster of KRAB-ZFPs that are located on chromosome 19p13.2. Thus, we focused on this locus to ascertain if the expression of other members of this cluster was altered. We have identified several KRAB-ZFPs that are differentially expressed and are associated with changes in overall survival in patients. Since the gene ZNF844 is a paralog of ZNF433, it appears to be a strong candidate biomarker and prognostic factor for ccRCC. 305
BackgroundThe study of hyperoxia vs. hypoxia on tumor cell survival is often studied under experi... more BackgroundThe study of hyperoxia vs. hypoxia on tumor cell survival is often studied under experimental conditions that do not control for the shift in pH.MethodsHere, we examine the molecular genetic effects of (~91% O2 (hyperoxia) vs. ~2.5% O2 (hypoxia)) at a uniform pH ~7.2 using microarrays, HPLC, imaging probes and Western blot.ResultsHyperoxic and Hypoxic culture conditions for 24 hrs had no adverse effect on viability in malignant N2a cells, provided pH was neutral with ample glucose supply. The hyperoxic transcriptome showed elevation of mRNA of 42 mitochondrial genes, DNA/base excision repair, G protein signaling, protein synthesis, transcription and ubiquitin‐mediated proteolysis. The hypoxic transcriptome corresponded to major elevations in epigenetic (lysine (K)‐specific demethylases, H4k/d/j, Hist4h4, Hist2h3c1, Rara and REST corepressor 2) with greater expression of protein/mRNA for solute carrier family 16, member 3 MCT‐4 (lactate transporter). Blocking MCT‐4 (phloret...
cells at higher cocaine doses compared with the unchallenged cells. Since high concentrations of ... more cells at higher cocaine doses compared with the unchallenged cells. Since high concentrations of NO are associated with inflammation, the high levels of NO production observed in the present study suggested that cocaine may have potentiated the inflammatory response in the challenged C6 astroglia-like cells.
Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit breast carcinogen... more Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit breast carcinogenesis. CCL2/MCP-1 has been shown to play a significant role in breast cancer. This study explored DATS efficacy on triple-negative breast cancer (TNBC) cells. Materials and Methods: DATS efficacy on TNFα induced TNBC cells were examined via trypan blue exclusion test, wound-healing assay, human cytokine arrays, ELISA, and RT-PCR. Results: DATS significantly induced cell death and inhibited cell migration. Expression of CCL2/MCP-1, IL-6, PDGF-BB, NT-3, and GM-CSF in TNF-α-treated cells increased. However, DATS significantly decreased the expression of CCL2/MCP-1 in TNF-α-treated MDA-MB-231 but not in MDA-MB-468 cells. DATS significantly down-regulated mRNA expression of IKBKE and MAPK8 in both cell lines, indicating a possible effect in genes involved in the NF-ĸB and MAPK signaling. Conclusion: DATS may have a role in TNBC therapy and prevention by targeting CCL2.
Within the past three centuries, all-cause disease burden in developed countries has shifted from... more Within the past three centuries, all-cause disease burden in developed countries has shifted from infectious to non-communicable (NCD)/genetic based diseases including cardiovascular conditions, cancer, neuropsychiatric conditions, and diabetes. Factors accounting for this drift include discoveries in vaccination (e.g., tetanus, cholera, typhoid, plague, anthrax, and tuberculosis), antibiotics, advances in medical diagnostics, lasers, surgical techniques, and routine medicines to treat almost every type of systemic imbalance. Moreover, advances in public health, sanitation, food safety, and geriatric sciences are creating extended life expectancy, where age-related illnesses (osteoarthritis, back pain, neurodegenerative conditions) in addition to NCDs are plaguing an ever-growing elderly population. The age-related risk for these diseases is now worsened by aggregation of global industrial pollutants, where the World Health Organization (WHO) now uses the term “environmental burden ...
chronic inflammation associated with cancer is characterized by the production of different types... more chronic inflammation associated with cancer is characterized by the production of different types of chemokines and cytokines. in cancer, numerous signaling pathways upregulate the expression levels of several cytokines and evolve cells to the neoplastic state. Therefore, targeting these signaling pathways through the inhibition of distinctive gene expression is a primary target for cancer therapy. The present study investigated the anticancer effects of the natural polyphenol gossypol (GoSS) in triple-negative breast cancer (TnBc) cells, the most aggressive breast cancer type with poor prognosis. GoSS effects were examined in two TnBc cell lines: Mda-MB-231 (MM-231) and Mda-MB-468 (MM-468), representing caucasian americans (ca) and african americans (aa), respectively. The obtained ic 50 s revealed no significant difference between the two cell lines' response to the compound. However, the use of microarray assays for cytokine determination indicated the ability of GoSS to attenuate the expression levels of cancer-related cytokines in the two cell lines. although GoSS did not alter ccl2 expression in MM-468 cells, it was able to cause 30% inhibition in TnF-α-stimulated MM-231 cells. additionally, il-8 was not altered by GoSS treatment in MM-231 cells, while its expression was inhibited by 60% in TnF-α-activated MM-468 cells. eliSa assays supported the microarray data and indicated that ccl2 expression was inhibited by 40% in MM-231 cells, and il-8 expression was inhibited by 50% in MM-468 cells. Furthermore, in MM-231 cells, GoSS inhibited ccl2 release via the repression of iKBKe, CCL2 and MAPK1 gene expression. additionally, in MM-468 cells, the compound downregulated the release of il-8 through repressing IL-8, MAPK1, MAPK3, CCDC88A, STAT3 and PIK3CD gene expression. in conclusion, the data obtained in the present study indicate that the polyphenol compound GoSS may provide a valuable tool in TnBc therapy.
Methamphetamine (METH) is a powerfully addictive psychostimulant that has a pronounced effect on ... more Methamphetamine (METH) is a powerfully addictive psychostimulant that has a pronounced effect on the central nervous system (CNS). The present study aimed to assess METH toxicity in differentiated C6 astroglia-like cells through biochemical and toxicity markers with acute (1 h) and chronic (48 h) treatments. In the absence of external stimulants, cellular differentiation of neuronal morphology was achieved through reduced serum (2.5%) in the medium. The cells displayed branched neurite-like processes with extensive intercellular connections. Results indicated that acute METH treatment neither altered the cell morphology nor killed the cells, which echoed with lack of consequence on reactive oxygen species (ROS), nitric oxide (NO) or inhibition of any cell cycle phases except induction of cytoplasmic vacuoles. On the other hand, chronic treatment at 1 mM or above destroyed the neurite-like processors and decreased the cell viability that paralleled with increased levels of ROS, lipid...
The Research Centers in Minority Institutions (RCMI) program was established by the US Congress ... more The Research Centers in Minority Institutions (RCMI) program was established by the US Congress to support the development of biomedical research infrastructure at minority-serving institutions granting doctoral degrees in the health professions or in a health-related science. RCMI institutions also conduct research on diseases that disproportionately affect racial and ethnic minorities (ie, African Americans/Blacks, American Indians and Alaska Natives, Hispanics, Native Hawaiians and Other Pacific Islanders), those of low socioeconomic status, and rural persons. Quantitative metrics, including the numbers of doctoral science degrees granted to underrepresented students, NIH peer-reviewed research funding, peer-reviewed publications, and numbers of racial and ethnic minorities participating in sponsored research, demonstrate that RCMI grantee institutions have made substantial progress toward the intent of the Congressional legislation, as well as the NIH/NIMHD-linked goals o...
Background/Aim: Chemotherapy resistance in triple-negative breast cancer (TNBC) cells is well doc... more Background/Aim: Chemotherapy resistance in triple-negative breast cancer (TNBC) cells is well documented. Therefore, it is necessary to develop safer and more effective therapeutic agents to enhance the outcomes of chemotherapeutic agents. The natural alkaloid sanguinarine (SANG) has demonstrated therapeutic synergy when coupled with chemotherapeutic agents. SANG can also induce cell cycle arrest and trigger apoptosis in various cancer cells. Materials and Methods: In this study, we investigated the molecular mechanism underlying SANG activity in MDA-MB-231 and MDA-MB-468 cells as two genetically different models of TNBC. We employed various assays including Alamar Blue to measure the effect of SANG on cell viability and proliferation rate, flow cytometry analysis to study the potential of the compound to induce apoptosis and cell cycle arrest, quantitative qRT PCR apoptosis array to measure the expression of different genes mediating apoptosis, and the western system was used to analyze the impact of the compound on AKT protein expression. Results: SANG lowered cell viability and disrupted cell cycle progression in both cell lines. Furthermore, S-phase cell cycle arrest-mediated apoptosis was found to be the primary contributor to cell growth inhibition in MDA-MB-231 cells. SANG-treated TNBC cells showed significantly up-regulated mRNA expression of 18 genes associated with apoptosis, including eight TNF receptor superfamily (TNFRSF), three members of the BCL2 family, and two members of the caspase (CASP) family in MDA-MB-468 cells. In MDA-MB-231 cells, two members of the TNF superfamily and four members of the BCL2 family were affected. The western study data showed the inhibition of AKT protein expression in both cell lines concurrent with up-regulated BCL2L11 gene. Our results point to the AKT/PI3K signaling pathway as one of the key mechanisms behind SANG-induced cell cycle arrest and death. Conclusion: SANG shows anticancer properties and apoptosis-related gene expression changes in the two TNBC cell lines and suggests AKT/PI3K pathway implication in apoptosis induction and cell cycle arrest. Thus, we propose SANG's potential as a solitary or supplementary treatment agent against TNBC. In the United States, breast cancer (BC) is the most common malignancy and the second leading cause of death among women aged 20-59 years (1). Gene expression is controlled by molecular signal transduction, and alterations in transcriptional settings distinguish cancer cells (2). BC is a heterogeneous cancer type categorized based on its molecular features. Triple-negative breast cancer (TNBC) affects approximately 15% of patients with BC (2). However, the incidence of this subtype is two to three folds higher in African American (AA) women than in other ethnic groups (3, 4). TNBC is the most aggressive BC and has a poor outcome compared to different BC subtypes (3, 5). Based on immunohistochemical characteristics, the expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) are missing in TNBC cells (3, 6). Currently, chemotherapy is the principal treatment for patients with TNBC. However, disease recurrence can manifest within the first two years 323
International Journal of Environmental Research and Public Health
Being cognizant of the pronounced health advantages of breastfeeding for both the nursing mother ... more Being cognizant of the pronounced health advantages of breastfeeding for both the nursing mother and her infant, the breastfeeding dyad, we examined breastfeeding rates among Floridian women who gave birth from 2012 to 2014 (N = 639,052). We investigated the associations between breastfeeding initiation and WIC-based breastfeeding support (the Special Supplemental Nutrition Program for Women, Infants, and Children), education level, and race and ethnicity. We compared the percentage of breastfeeding mothers between those in the WIC program to those who were not, and we compared breastfeeding rates across racial and ethnic groups. Consistent with previous reports, black newborns in this study were breastfed at lower rates than other racial groups, and WIC program participants were less likely to breastfeed than non-WIC program participants. However, by breaking down the data by education level and race, and ethnicity, we see a significantly increased rate of breastfeeding due to WIC ...
Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit carcinogenesis in... more Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit carcinogenesis in cancer cells. We have previously shown DATS's ability to decrease the percentage of viable cells, inhibit cell migration and modulate genes involved in the nuclear factor kappa-light-chainenhancer of activated B cells (NF-ĸB) and mitogen-activated protein kinase (MAPK) signaling. Materials and Methods: This study aimed to compare the efficacy of DATS in tumor necrosis factor alpha (TNF-α) induced MDA-MB-231 and MDA-MB-468 cells and investigate its role in cell-death signaling via cell cycle, flow cytometry, and caspase assay. Results: DATS exhibit a time-dependent accumulation of G 2 /M phase cells in both cell lines, with higher effects in the MDA-MB-468 for all time points. DATS's ability to decrease the percentage of viable cells in both MDA-MB-231 and MDA-MB-468 cells was shown by a significant but slight increase of early and late apoptosis in the presence of DATS compared to control. Moreover, MDA-MB-468 cells showed more sensitivity to the DATS effect, evidenced by the higher percentage of apoptosis than MDA-MB-231 cells. The caspase studies showed a significant increase in caspase 3 and 8 activity in the presence of DATS, compared to control, in both cell lines. DATS showed no significant increase in caspase 9 activity in both cell lines compared to the control. Conclusion: DATS-induced apoptosis in human breast cancer cells is mediated, at least in part, by cell cycle arrest and caspase activity. These findings provide information for future studies into the role of DATS in TNBC therapy and prevention.
ASPET 2023 Annual Meeting Abstract - Drug Discovery and Development
ID 14491 Poster Board 296 For thousands of years, humans have been orally consuming herbal botani... more ID 14491 Poster Board 296 For thousands of years, humans have been orally consuming herbal botanical medicines, ascribing the beneficial effects to the plant itself. Oddly, we have not yet realized the impact of ingesting dead/live microorganisms in these plants. The residual dead remains of the herbal microbiome can and do leave a trail of thermal and pH-stable immune-stimulating antigenic epitopes with the capacity to act in the human gut. The current study employs a high throughput screening to elucidate the immune-stimulating properties of over-the-counter (OTC) / point-of-sale (POS) herbs. The findings illustrate the existence of an immune active herbal bioactive microbiome, which is plant specific but independent of the plant itself. In this study, an initial high throughput screening of 2535 phytochemicals and OTC/POS'S botanicals was carried out to elucidate pro-inflammatory effects in RAW 264.7 macrophages, with many as potent as positive controls: lipopolysaccharide (LPS) (E.coli 0111:B4). Microbial pathogen-associated molecular pattern molecules (PAMPS) in bio-active herbs were removed from the plants by spin column application, extracts were retested for a bioactive response, and the pH stability of LPS analyzed using a stomach simulation model. Lastly, we reverse cultured the "residual remains" in 7 OTC/POS herbs which were isolated, colonized, and sequenced using Illumina targeting the V4 region of the 16S rRNA gene and the V1-V3 region of the 18S rRNA gene. The obtained results show that Out of 2535 compounds screened, not a single phytochemical or drug (>500 tested) could mimic LPS, which was only observed for 65 "specific" herbal medicine plants, having high concentrations of PAMPS, dominating in roots and oceanic marine sea kelps /algae's [e.g.
ASPET 2023 Annual Meeting Abstract - Cancer Pharmacology
ID 54503 Poster Board 446 Triple-negative breast cancer (TNBC) has historically had fewer treatme... more ID 54503 Poster Board 446 Triple-negative breast cancer (TNBC) has historically had fewer treatment alternatives than other breast cancer types. Therefore, it is critical to explore and pinpoint potential biomarkers that could be utilized in treating TNBC. By doing this, we will significantly enhance a patient's prognosis and quality of life. Apoptosis-controlling genes could be manipulated to increase the death of cancer cells. It is essential to comprehend how the genes involved in the apoptotic pathway interact to identify potential therapeutic targets. Therefore, the current study is aimed to evaluate the gene expression, protein-protein interaction (PPI), and transcription factor interaction of 27 apoptosis-regulated genes in TNBC using integrated bioinformatics methods to assess the gene expression, protein-protein interaction (PPI) and transcription factor interaction. Our findings demonstrated that CASP2, CASP3, DAPK1, TNF, TRAF2, and TRAF3 were substantially associated with the overall survival rate (OS) difference of TNBC patients. The results also show that breast tissue gene expressions of BNIP3, TNFRSF10B, MCL1, and CASP4 were downregulated compared to normal breast tissue in UALCAN. At the same time, BIK, AKT1, BAD, FADD, DIABLO, and CASP9 were downregulated in bc-GeneEx-Miner v4.5 mRNA expression (BCGM) databases. Based on the GO term enrichment analysis, the Cellular process (GO:0009987), which has about 21 apoptosis-regulated genes, is the top category in the biological process (BP), followed by biological regulation (GO:0065007). We identified 29 pathways, including p53, angiogenesis, and apoptosis signaling. We examined the PPIs between the genes that regulate apoptosis; CASP3 and CASP9 interact with FADD, MCL1, TNF, TNFRSRF10A, and TNFRSF10; additionally, CASP3 significantly forms PPIs with CASP9, DFFA, and TP53, while CASP9 with DIABLO. In the top 10 transcription factors, the androgen receptor (AR) interacts with five apoptosis-regulated genes (p<0.0001; q<0.01), followed by RARA (Retinoic Acid Receptor Alpha) (p<0.0001; q<0.01) and RNF2 (Ring Finger Protein) (p<0.0001; q<0.01). Overall, the gene expression profile, the PPIs, and the apoptosis-TF interaction findings suggested that the 27 apoptosis-regulated genes might be used as promising targets in treating and managing TNBC. Furthermore, from a total of 27 key genes, CASP2, CASP3, DAPK1, TNF, TRAF2, and TRAF3 were significantly correlated with poor overall survival in TNBC (p-value < 0.05), which could play essential roles in the progression of TNBC and provide attractive therapeutic targets that may offer new candidate molecules for targeted therapy. It was concluded from this study that genetic alterations altering apoptosis could be possible biomarkers for the prognosis and facilitate the search for alternative treatment targets for TNBC.
Muscadine grapes accumulate higher amounts of bioactive phenolics compared with other grape speci... more Muscadine grapes accumulate higher amounts of bioactive phenolics compared with other grape species. To identify the molecular events associated with polyphenolic accumulation that influence antioxidant capacity, two contrasting muscadine genotypes (C5 and C6) with varied phenolic/flavonoid content and antioxidant activity were investigatedviaRNA-sequencing during berry development. The results showed that berry development is concomitant with transcriptome profile changes, which was more pronounced at the véraison (V) stage. Despite that the downregulation pattern of gene expression dominated the upregulation through berry development, the C5 genotype maintained higher expression levels. Comparative transcript profiling allowed the identification of 94 differentially expressed genes with potential relevance in regulating fruit secondary metabolism, including 18 transcription factors and 76 structural genes. The genes underlying the critical enzymes in the modification reactions of ...
Muscadine berries display enhanced nutraceutical value due to the accumulation of distinctive phy... more Muscadine berries display enhanced nutraceutical value due to the accumulation of distinctive phytochemical constituents with great potential antioxidant activity. Such nutritional and health merits are not only restricted to muscadine, but muscadine berries accumulate higher amounts of bioactive polyphenolics compared with other grape species. For the genetic study of the antioxidant trait in muscadine, a multi-locus genome-wide association study (GWAS) with 350 muscadine genotypes and 1,283 RNase H2 enzyme-dependent amplicon sequencing (rhAmpSeq) markers was performed. Phenotyping was conducted with several antioxidant-related traits, including total phenolic content (TPC), total flavonoid content (TFC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, and FRAP antioxidant assay in muscadine berry skin. The correlation coefficient analysis revealed that the TPC, and DPPH/FRAP activities were significantly correlated. Through the GWAS analysis, 12 QTNs were id...
A distinctive tumor microenvironment associated with chronic inflammation, cancer development, an... more A distinctive tumor microenvironment associated with chronic inflammation, cancer development, and drug resistance in triple-negative breast cancer (TNBC) affect disproportionally Caucasian and African American women. PURPOSE: This investigation presents possible new biomarkers that could be modulated by the natural compound pentagalloyl glucose (PGG) in MM-231 (Caucasians) and MM-468 (African American) TNBC cells. METHODS: Cytokine arrays, ELISA, RT-PCR, flow cytometry, and Western analysis were performed. RESULTS: PGG reduced the expression of GRO-α/CXCL1 pro-inflammatory cytokine, which is associated with tumor recurrence and chemoresistance. PGG inhibited IκBKE and MAPK1 gene and protein expression indicating a possible signaling pathway for CXCL1 downregulation through NFκB and MAPK signaling. PGG also inhibited cell growth in both cell lines, which could be associated with a decrease in CXCL1 levels. Besides, PGG induced apoptosis, modulating numerous genes, including caspases...
Rosmarinic acid (RA) is a polyphenolic compound with various pharmacological properties, includin... more Rosmarinic acid (RA) is a polyphenolic compound with various pharmacological properties, including anti-inflammatory, immunomodulatory, anti-mutagenic, antioxidant, and anticancer activities. This study evaluated the effects of RA in two racially different triple-negative breast cancer (TNBC) cell lines. One cell line was derived from Caucasian origin (MDA-MB-231), and another one was derived from an African American origin (MDA-MB-468). The apoptotic effect of RA was performed using An Annexin V-FITC Apoptosis Detection Kit, while the Propidium Iodide Flow Cytometry Kit was used to detect the cell cycle distribution. Gene expression profiling was established using Human Apoptosis PCR array. The results obtained show that RA significantly caused cytotoxic and antiproliferative effects in both cell lines in a dose- and time-dependent manner. It was noted that RA induced cell cycle arrest-related apoptosis and altered the expression of many apoptosis-involved genes differently. In MDA...
Background/Aim: Clear-cell renal cell carcinoma (ccRCC) is the most common and aggressive form of... more Background/Aim: Clear-cell renal cell carcinoma (ccRCC) is the most common and aggressive form of all urological cancers, with poor prognosis and high mortality. Despite growing evidence of involvement in carcinogenesis, the role of KRAB-ZFP in ccRCC has not been fully explored. KRAB Zinc finger proteins (KRAB-ZFPs) are the largest family of mammalian transcription regulators. They are differentially expressed in various tissues during cellular development and phenotypic differentiation. Materials and Methods: In this study, the levels of transcripts of ccRCC from The Cancer Genome Atlas (TCGA) dataset were used to identify prognostic biomarkers in this disease. Results: Using bioinformatics techniques, we demonstrate that approximately 60% of KRAB zinc finger proteins located on chromosome 19p13.2 are differentially expressed, with all but two being down-regulated in ccRCC. Moreover, ZNF844, a paralog of ZNF433, was the most down-regulated across all histological grades and pathological stages (p<0.001). In addition, the decrease in ZNF844 expression was associated with poor patient survival (HR=0.41; 95% CI=0.3-0.56; p<0.0001). Gene Set Enrichment Analysis of genes inversely co-expressed with ZNF844 revealed that enriched pathways were consistently related to immune and translation processes (p<0.05, FDR <0.05). Lastly, ZNF844 expression showed moderate, inverse correlation to Helper Tcell (CD4 or Th1) subtype 1 (R=-0.558, p=5.15×10-39) infiltration and with the exhausted T-cell phenotype (R=-0.37; p=4.1×10-21). Conclusion: Down-regulation of KRAB-ZFPs at 19p13.2 may represent a signature for ccRCC. Moreover, ZNF844 is a prognostic marker for ccRCC and may serve as a putative immune-related tumor suppressor gene. Clear cell renal carcinoma (ccRCC) is the most common neoplastic disease affecting the kidneys. It accounts for at least 70% of all kidney cancers (1, 2). In advanced stages, ccRCC becomes resistant to conventional therapies (1-3). Moreover, later stages are associated with poor prognosis with a 5-year overall survival rate of less than 10% (1-3). Despite growing evidence of involvement in carcinogenesis, the role of KRAB-ZFP in ccRCC has not been fully explored. Kruppel-associated box domain-containing zinc-finger proteins (KRAB-ZFP) represent the most prominent family of transcriptional repressors (4, 5). The human genome contains over 300 KRAB-ZFPs genes that produce more than 800 transcripts that encode proteins, pseudogenes, and splice variants (6-8). KRAB-ZFPs are differentially expressed in various tissues during cellular development. Several of these genes are involved in embryogenesis, genome imprinting, and biological processes that determine phenotypes in stem cells (9-11). Given their roles in cellular differentiation and their ability to alter gene expression via epigenetic processes and chromatin remodeling, it is conceivable that dysregulation of KRAB-ZFP transcription factors could facilitate tumorigenesis and tumor progression. Interestingly, several large KRAB-ZFP clusters are located on chromosome 19 (chr19), and several studies have linked genetic alterations in chromosome 19p13 to cancers (12-18). Using data mining techniques, we recently identified ZNF433, a zinc finger that is differentially expressed in ccRCC (19) and that may be a biomarker for ccRCC. ZNF433 is part of a cluster of KRAB-ZFPs that are located on chromosome 19p13.2. Thus, we focused on this locus to ascertain if the expression of other members of this cluster was altered. We have identified several KRAB-ZFPs that are differentially expressed and are associated with changes in overall survival in patients. Since the gene ZNF844 is a paralog of ZNF433, it appears to be a strong candidate biomarker and prognostic factor for ccRCC. 305
BackgroundThe study of hyperoxia vs. hypoxia on tumor cell survival is often studied under experi... more BackgroundThe study of hyperoxia vs. hypoxia on tumor cell survival is often studied under experimental conditions that do not control for the shift in pH.MethodsHere, we examine the molecular genetic effects of (~91% O2 (hyperoxia) vs. ~2.5% O2 (hypoxia)) at a uniform pH ~7.2 using microarrays, HPLC, imaging probes and Western blot.ResultsHyperoxic and Hypoxic culture conditions for 24 hrs had no adverse effect on viability in malignant N2a cells, provided pH was neutral with ample glucose supply. The hyperoxic transcriptome showed elevation of mRNA of 42 mitochondrial genes, DNA/base excision repair, G protein signaling, protein synthesis, transcription and ubiquitin‐mediated proteolysis. The hypoxic transcriptome corresponded to major elevations in epigenetic (lysine (K)‐specific demethylases, H4k/d/j, Hist4h4, Hist2h3c1, Rara and REST corepressor 2) with greater expression of protein/mRNA for solute carrier family 16, member 3 MCT‐4 (lactate transporter). Blocking MCT‐4 (phloret...
cells at higher cocaine doses compared with the unchallenged cells. Since high concentrations of ... more cells at higher cocaine doses compared with the unchallenged cells. Since high concentrations of NO are associated with inflammation, the high levels of NO production observed in the present study suggested that cocaine may have potentiated the inflammatory response in the challenged C6 astroglia-like cells.
Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit breast carcinogen... more Background/Aim: Diallyl trisulfide (DATS) has been shown to prevent and inhibit breast carcinogenesis. CCL2/MCP-1 has been shown to play a significant role in breast cancer. This study explored DATS efficacy on triple-negative breast cancer (TNBC) cells. Materials and Methods: DATS efficacy on TNFα induced TNBC cells were examined via trypan blue exclusion test, wound-healing assay, human cytokine arrays, ELISA, and RT-PCR. Results: DATS significantly induced cell death and inhibited cell migration. Expression of CCL2/MCP-1, IL-6, PDGF-BB, NT-3, and GM-CSF in TNF-α-treated cells increased. However, DATS significantly decreased the expression of CCL2/MCP-1 in TNF-α-treated MDA-MB-231 but not in MDA-MB-468 cells. DATS significantly down-regulated mRNA expression of IKBKE and MAPK8 in both cell lines, indicating a possible effect in genes involved in the NF-ĸB and MAPK signaling. Conclusion: DATS may have a role in TNBC therapy and prevention by targeting CCL2.
Within the past three centuries, all-cause disease burden in developed countries has shifted from... more Within the past three centuries, all-cause disease burden in developed countries has shifted from infectious to non-communicable (NCD)/genetic based diseases including cardiovascular conditions, cancer, neuropsychiatric conditions, and diabetes. Factors accounting for this drift include discoveries in vaccination (e.g., tetanus, cholera, typhoid, plague, anthrax, and tuberculosis), antibiotics, advances in medical diagnostics, lasers, surgical techniques, and routine medicines to treat almost every type of systemic imbalance. Moreover, advances in public health, sanitation, food safety, and geriatric sciences are creating extended life expectancy, where age-related illnesses (osteoarthritis, back pain, neurodegenerative conditions) in addition to NCDs are plaguing an ever-growing elderly population. The age-related risk for these diseases is now worsened by aggregation of global industrial pollutants, where the World Health Organization (WHO) now uses the term “environmental burden ...
chronic inflammation associated with cancer is characterized by the production of different types... more chronic inflammation associated with cancer is characterized by the production of different types of chemokines and cytokines. in cancer, numerous signaling pathways upregulate the expression levels of several cytokines and evolve cells to the neoplastic state. Therefore, targeting these signaling pathways through the inhibition of distinctive gene expression is a primary target for cancer therapy. The present study investigated the anticancer effects of the natural polyphenol gossypol (GoSS) in triple-negative breast cancer (TnBc) cells, the most aggressive breast cancer type with poor prognosis. GoSS effects were examined in two TnBc cell lines: Mda-MB-231 (MM-231) and Mda-MB-468 (MM-468), representing caucasian americans (ca) and african americans (aa), respectively. The obtained ic 50 s revealed no significant difference between the two cell lines' response to the compound. However, the use of microarray assays for cytokine determination indicated the ability of GoSS to attenuate the expression levels of cancer-related cytokines in the two cell lines. although GoSS did not alter ccl2 expression in MM-468 cells, it was able to cause 30% inhibition in TnF-α-stimulated MM-231 cells. additionally, il-8 was not altered by GoSS treatment in MM-231 cells, while its expression was inhibited by 60% in TnF-α-activated MM-468 cells. eliSa assays supported the microarray data and indicated that ccl2 expression was inhibited by 40% in MM-231 cells, and il-8 expression was inhibited by 50% in MM-468 cells. Furthermore, in MM-231 cells, GoSS inhibited ccl2 release via the repression of iKBKe, CCL2 and MAPK1 gene expression. additionally, in MM-468 cells, the compound downregulated the release of il-8 through repressing IL-8, MAPK1, MAPK3, CCDC88A, STAT3 and PIK3CD gene expression. in conclusion, the data obtained in the present study indicate that the polyphenol compound GoSS may provide a valuable tool in TnBc therapy.
Methamphetamine (METH) is a powerfully addictive psychostimulant that has a pronounced effect on ... more Methamphetamine (METH) is a powerfully addictive psychostimulant that has a pronounced effect on the central nervous system (CNS). The present study aimed to assess METH toxicity in differentiated C6 astroglia-like cells through biochemical and toxicity markers with acute (1 h) and chronic (48 h) treatments. In the absence of external stimulants, cellular differentiation of neuronal morphology was achieved through reduced serum (2.5%) in the medium. The cells displayed branched neurite-like processes with extensive intercellular connections. Results indicated that acute METH treatment neither altered the cell morphology nor killed the cells, which echoed with lack of consequence on reactive oxygen species (ROS), nitric oxide (NO) or inhibition of any cell cycle phases except induction of cytoplasmic vacuoles. On the other hand, chronic treatment at 1 mM or above destroyed the neurite-like processors and decreased the cell viability that paralleled with increased levels of ROS, lipid...
The Research Centers in Minority Institutions (RCMI) program was established by the US Congress ... more The Research Centers in Minority Institutions (RCMI) program was established by the US Congress to support the development of biomedical research infrastructure at minority-serving institutions granting doctoral degrees in the health professions or in a health-related science. RCMI institutions also conduct research on diseases that disproportionately affect racial and ethnic minorities (ie, African Americans/Blacks, American Indians and Alaska Natives, Hispanics, Native Hawaiians and Other Pacific Islanders), those of low socioeconomic status, and rural persons. Quantitative metrics, including the numbers of doctoral science degrees granted to underrepresented students, NIH peer-reviewed research funding, peer-reviewed publications, and numbers of racial and ethnic minorities participating in sponsored research, demonstrate that RCMI grantee institutions have made substantial progress toward the intent of the Congressional legislation, as well as the NIH/NIMHD-linked goals o...
Uploads
Papers by Karam Soliman