Breast cancer (BC) is frequent among women in worldwide as well as in India. Several studies have... more Breast cancer (BC) is frequent among women in worldwide as well as in India. Several studies have reported a wide variation (1.6-86.2%) in the frequency of incidence of human papillomavirus (HPV) infection in BC with high prevalence of high risk HPV16 subtype. HPV infection in breast can occur through different routes like body fluid or by micro-lesion of breast skin from genital/agential sites, though the actual mode of HPV transmission is not yet known in details. Frequent integration and sequence variation with low copy number of HPV16 were seen in this tumour. In addition, high frequencies of methylation in p97 promoter region of HPV16 were evident in this tumour. Novel splice variants of E6/E7 along with other common variants and their protein expression were seen in the tumour. This indicates the importance of HPV in this tumor, its early diagnosis and prognosis. Thus, HPV may be targeted through vaccination to control the disease. However, detailed analysis of HPV associated molecular pathogenesis of BC is warranted for proper therapeutic intervention.
Indian Journal of Medical and Paediatric Oncology, Apr 1, 2021
Introduction The improved prognosis of pediatric B-cell acute lymphoblastic leukemia (pBALL) is c... more Introduction The improved prognosis of pediatric B-cell acute lymphoblastic leukemia (pBALL) is considered as a good progress of medical science in the field of oncology and hematology. Minimal residual disease (MRD) refers to presence of disease in molecular level is a newer practice with respect to the detection of complete remission by conventional pathologic analysis. Prognostic value of MRD in pediatric ALL (p-ALL) is well known. Objectives This study was aimed to describe clinical outcomes and prognosis, that is, overall survival and relapse in the patients with pBALL with respect to minimal residual disease detection on day 15, day 29, and postconsolidations in a tertiary care center in eastern India. Materials and Methods Eight color flow cytometry was used to detect MRD in this study. This contained markers such as CD 19, CD 34, CD 10, CD58, CD 45, CD13, anti-TDT, CD33. Eight panels included were (1) CMPO-FITC/cCD79a-PE/cCd3ECD, (2) CD20-FITC/cCD10-PE/cCd-19ECD, (3) CD34-FITC/cCD117-PE/cCd45 ECD/CD2 PC 5, (4) CD15 FITC/ CD33PE/CD45ECD, (5) CD14 FITC/CD13 PE/CD45ECD, (6) HLADR FITC/ CD7 PE/CD45 ECD, (7) TdT FITC/CD45 ECD (IF CD34 NEG), and (8) CD58 FITC/CD 45 ECD (IF BOTH CD34 AND TdT NEG; were used to prepare the marker. Results The study included 52 patients. In the 52 patients, 59.6% patients are alive with a p-value of 0.031. MRD was checked on every 15th and the 29th day and postconsolidation of the treatment where in day 15 (p = 0.023), it was 53.4% positive and 46.5% negative. On day 29 (p = 0.031), MRD was 22.5% positive and 77.5% negative, in post consolidation, it was positive in 20% and negative is 80%. MRD value below 0.01 is taken as negative and above is taken as positive. The overall survival (OS) is of 32.88 + 8.59 with a 6 to 36 months of duration. In relapsed cases, no hemorrhagic relapse was found and two CNS relapses were found. Conclusion It was a study of 52 patients of pBALL with a detection of MRD by FCM. MRD-negative patients had a good prognosis and comparatively lower rate of relapse than the one with positive MRD. Effort should be made to adhere to recommendation of MRD testing in clinical guidelines.
Mouse plasmacytomas (MPCs) induced by pristane oil, or by a combination of pristane oil and Abels... more Mouse plasmacytomas (MPCs) induced by pristane oil, or by a combination of pristane oil and Abelson virus, carry one of two chromosomal translocations. The typical 12; 15 translocation leads to the juxtaposition of c-myc and immunoglobulin heavy-chain sequences, whereas the 6; 15 translocation links the kappa light-chain locus with the pvt-1 (plasmacytoma variant translocation) locus, located at least 75kb 3' of c-myc [Cory, S., Graham, M., Webb, E., Corcoran, L. & Adams, J. (1985). EMBO J., 4, 675-681]. Unlike the human Burkitt's lymphoma-associated translocation, the lambda/myc juxtaposed variant translocation has not been found previously in MPCs. Using unconventional MPC induction systems in which the tumor precursor cell was induced to proliferate in a secondary host, we have recently identified a 15; 16 translocation in six of the derived MPCs [Wiener, F., Silva, S., Sugiyama, H., Babonits, M. & Klein, G. (1990). Genes Chromosomes Cancer, 2, 36-43]. Chromosome 16 harbors the lambda light-chain gene. To explore whether the 15; 16 translocation represents the lambda/myc juxtaposition, we have mapped the breakpoints on chromosomes 15 and 16 by pulsed-field gel electrophoresis (PFGE). The pvt-1 region was mapped to approximately 220 kb 3' of c-myc. The breakpoint on chromosome 15 in ABPC-Ch-163-10, one of the six 15; 16 translocation-carrying MPCs, was situated approximately 80 kb 3' of c-myc and 140 kb 5' of pvt-1b, the major breakpoint cluster region of the previously analysed 6; 15 variant MPCs. The breakpoint on chromosome 16 was found to cut between the V1 and C3 regions of the lambda locus. Co-migration experiments showed that the C3 and the myc gene were juxtaposed head to tail on the 15; 16 translocation chromosome. On the reciprocal product V1 was juxtaposed to pvt-1.
Background & objectives: Medicinal plants like Swertia chirata are rich sources of different ... more Background & objectives: Medicinal plants like Swertia chirata are rich sources of different xanthones. This study was aimed to assess the cytotoxic potential of four most abundant xanthones present in S. chirata both in vivo and in vitro in Ehrlich ascites carcinoma (EAC), a mouse transplantable breast carcinoma cell line and two human breast carcinoma cell lines (MCF-7 and MDA-MB-231). Methods: Four xanthones derived from S. chirata namely 1-hydroxy-3,7,8-trimethoxyxanthone (XA), 1,8-dihydroxy-3,5-dimethoxyxanthone (XB), 1-hydroxy-3,5,8-trimethoxyxanthone (XC) and 1,5,8-trihydroxy-3-methoxyxanthone (XD) were used for determination of sub-lethal dose on the cell lines EAC, MCF-7, MDA-MB-231 and verified toxicity of sub-lethal dose on normal murine fibroblast cells. Cytotoxicity was measured in vivo and survivability of mice was plotted accordingly. Therapeutic efficacy of XD was evaluated both in vivo and in vitro by determination of lipid peroxidation (LPO), reactive oxygen species (ROS) generation and by quantitating the enzyme status (GSH, catalase, superoxide dismutase) in treated and untreated samples. DNA damage was evaluated using comet and DNA fragmentation assays. Furthermore, apoptotic effect was analyzed by flow cytometry and validated by TUNEL assay and Western blotting. Results: Among all the xanthones tested XD showed IC50at the lowest dose, and normal cells were unaffected at this dose. Survivability of mice increased significantly when treated with XD compared to other xanthones and cisplatin. Significantly increased ROS and LPO were found in cancer cells as a result of XD treatment which was unaltered in normal cell line. XD induced DNA damage and apoptosis in cancer cell lines. Interpretation & conclusions: Our experimental data indicate that XD may potentially act as a chemotherapeutic agent by enhancing ROS in breast cancer cells thereby leading to apoptosis.
The present study was designed to determine the chemopreventive efficiency of 1,5,8-trihydroxy-3-... more The present study was designed to determine the chemopreventive efficiency of 1,5,8-trihydroxy-3-methoxy xanthone, abbreviated as TMX, isolated and purified from the aerial part of the plant Swertia chirata against 9,10-dimethylbenz[a]-anthracene (DMBA)/croton oilinduced skin cancer, and probe into the molecular mechanism. All the mice in the carcinogen control group developed severe dysplastic lesions after the 14th week of application of the carcinogen, which progressed to carcinoma in situ around the 20th week; this was validated histologically. However, after TMX treatment, only around 50% of mice developed papilloma which histologically was found to be restrictive to moderate to severe hyperplastic change in the 14th and 20th week. The chemopreventive potential was determined by calculating the attributable risk (AR), which was-11.3 for the 14th week and increased up to-17.5 for the 20th week. To ascertain the effect of TMX treatment on inflammation, the effect of TMX on inflammatory cytokines was studied by ELISA. It revealed a significant reduction in inflammation upon TMX treatment for the 20th week. As TMX could hold its chemopreventive potential up to the 20th week, the molecular mechanism of restriction was studied for the 20th week of treatment. Skin forms a rich source of stem cells which orchestrate the progression of carcinogenesis and become cancer stem cells (CSCs). β-Catenin and KRAS are known central modulators of CSCs, that play a crucial role in the progression of skin carcinogenesis. We observed that TMX treatment inhibited KRAS and nuclear translocation of β-catenin causing its cytoplasmic degradation by P-53 and P-21mediated pathway, thereby exerting its chemopreventive potential.
Head and neck squamous cell carcinoma is one of the leading cancers in terms of incidence and mor... more Head and neck squamous cell carcinoma is one of the leading cancers in terms of incidence and mortality. However, no reliable marker till date accurately predicts its progression when altered in healthy tissues. The study aims to identify alleles of microsatellites adjacent to important cell cycle regulatory, tumor suppressor genes altered in early head and neck lesions, viz. RBSP3, LIMD1 and CDC25A, which undergo frequent deletion and can be used for population screening and early detection. DNA for tumors and normal tissues was isolated from 143 patients in different stages of head and neck squamous cell carcinoma. The size of microsatellite present in normal tissues and their deletion in the corresponding tumor was identified, along with the correlation of expression in normal epithelium with respect to allele size. The results revealed a range of alleles (CA 9 to CA 32) for the different microsatellites of the genes in normal tissues. The larger alleles were significantly deleted with differential deletion of alleles observed in tumors, except for LIMD1, in which the smaller allele was significantly deleted. In normal tissues, some alleles represented as stable alleles with high prevalence, while in tumours, specific sizes showed greater propensity for deletion. However, similar expression of the proteins in normal epithelium adjacent to tumors was observed despite variations in allele size, possibly due to the location of the microsatellites. Thus, those alleles when present in normal tissues and undergoing persistent deletion in tumours could be used as markers for screening and early identification of populations at risk of developing head and neck lesions.
Background and Objectives: The aim of the study was to understand the importance of mismatch repa... more Background and Objectives: The aim of the study was to understand the importance of mismatch repair genes MLH1 and MSH2 in chemotolerance and prognosis of breast carcinoma (BC). Methods: First, the alterations (deletion/methylation/expression) of MLH1 and MSH2 were analyzed in 45 neoadjuvant chemotherapy (NACT)-treated and 133 pretherapeutic BC samples. The chemotolerant BC cells were characterized by treating two BC cell lines MCF-7 and MDA MB 231 with two anthracycline antitumor antibiotics, doxorubicin and nogalamycin. Results: The deletion frequencies were 32% to 38% in MLH1/MSH2 genes and promoter methylation frequencies were 49% to 62% in MLH1 and 41% to 51% in MSH2 in both NACT-treated and pretherapeutic samples. The overall alteration of MLH1 and MSH2 was 58% to 71% in the samples. Reduced messenger RNA (mRNA) and protein expression were found in both the genes and it showed concordance with the molecular alterations. NACT-treated patients showed better prognosis. The chemotherapeutic drug induced increased mRNA/protein expression of the genes in BC cell lines was due to their promoter hypomethylation, as analyzed by quantitative methylation assay. This phenomenon was also evident in NACT-treated BC samples. Conclusion: MLH1/MSH2 genes play a critical role in the development of BC. Hypomethylation of MLH1/MSH2 genes might be important in chemotolerance of the disease.
Background: Genetic instability of chromosome 11 is a frequent event in many solid tumours, inclu... more Background: Genetic instability of chromosome 11 is a frequent event in many solid tumours, including head and neck squamous cell carcinoma (HNSCC). Aims: To perform allelic imbalance analysis of cytogenetically mapped altered regions of human chromosome 11 in patients with HNSCC from eastern India. Methods: Genomic alterations were investigated using highly polymorphic microsatellite markers in both HNSCC and leukoplakia tissues. Results: Microsatellite markers D11S1758 from 11p13-15 and D11S925 from 11q23.3-24 had the highest frequency (38% and 32%, respectively) of loss of heterozygosity among all the markers analysed. Allelic loss at the marker D11S925 was seen in both leukoplakia and in all stages of HNSCC tumour tissues suggesting that it is an early event in HNSCC tumorigenesis. Microsatellite size alteration was also found to be high (> 20%) in several markers. In leukoplakia samples microsatellite instability was seen at a higher frequency than loss of the allele, indicating such alterations might initiate the process of tumorigenesis in HNSCC. Conclusions: The high rate of chromosomal alterations at 11q21-24 in HNSCC suggests the presence of a putative tumour suppressor gene in this region.
Purpose In approximately 30% of triple-negative breast cancer (TNBC) patients a complete patholog... more Purpose In approximately 30% of triple-negative breast cancer (TNBC) patients a complete pathological response is achieved. However, after neo-adjuvant chemotherapy treatment (NACT) residual tumour cells can be intrinsically resistant to chemotherapy. In this study, associations of the WNT/beta-catenin pathway with chemo-tolerance of NACT treated TNBC patients were compared to that of pre-treatment TNBC patients. Methods Expression analyses were performed in both pre-treatment and NACT treated TNBC samples using immunohistochemistry and qRT-PCR, along with DNA copy number variation (CNV) and promoter methylation analyses to elucidate the mechanism(s) underlying chemo-tolerance. In addition, in vitro validation experiments were performed in TNBC cells followed by in vivo clinicopathological correlation analyses. Results A reduced expression (41.1%) of nuclear beta-catenin together with a low proliferation index was observed in NACT samples, whereas a high expression (59.0%) was observed in pre-treatment samples. The reduced nuclear expression of beta-catenin in the NACT samples showed concordance with reduced expression levels (47-52.9%) of its associated receptors (FZD7 and LRP6) and increased expression levels (35.2-41.1%) of its antagonists (SFRP1, SFRP2, DKK1) compared to those in the pre-treatment samples. The expression levels of the receptors showed no concordance with its respective gene copy number/mRNA expression statuses, regardless treatment. Interestingly, however, significant increases in promoter hypomethylation of the antagonists were observed in the NACT samples compared to the pre-treatment samples. Similar expression patterns of the antagonists, receptors and beta-catenin were observed in the TNBC-derived cell line MDA-MB-231 using the anthracyclines doxorubicin and nogalamycin, suggesting the importance of promoter hypomethylation in chemotolerance. NACT patients showing reduced receptor and/or beta-catenin expression levels and high antagonist expression levels exhibited a comparatively better prognosis than the pre-treatment patients. Conclusions Our data suggest that reduced nuclear expression of beta-catenin in NACT TNBC samples, due to downregulation of its receptors and upregulation of its antagonists through promoter hypomethylation of the WNT pathway, plays an important role in chemo-tolerance.
Atlas of genetics and cytogenetics in oncology and haematology, Nov 1, 2011
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 ... more This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence.
Differential alterations of the spliceosomal Uridylic acid rich small nuclear RNAs (UsnRNAs) (U1,... more Differential alterations of the spliceosomal Uridylic acid rich small nuclear RNAs (UsnRNAs) (U1, U2, U4, U5, and U6) are reported to be associated with cellular proliferation and development, but definitive information is scarce and also elusive. An attempt is made in this study to analyze the metabolic patterns of major spliceosomal UsnRNAs, during tumor development, in an in vitro carcinogenesis model of 20-methylcholanthrene (MCA)-transformed Swiss Mouse Embryonic Fibroblast (MEF), designated as CNCI-PM-20. MEF cells, after treatment with 20-MCA, progressed through a sequence of passages with distinct and heritable changes, finally becoming neoplastic at passage-42 (P42). A differential expression pattern of major UsnRNAs was observed during this process. The abundance of U1 was 20% below control (P1) at passage-20 (P20), followed by a gradual increase up until P42 ($12% above the P1 value). The abundance of U2 was more or less constant during the cellular transformation. U4 showed a trend of increase, with above 30% abundance than control at P20, followed by a significant increase at P36 and P42 (1.5-and 2-fold, respectively, P-value <0.01). U5 also followed an identical pattern, with an increase of 70% compared to control (P-value <0.05) at P42. Interestingly, U6 gradually decreased from P20 onwards up until P42, with 22% at P20 and 67% at P42 (P-value <0.01). An overall significant quantitative alteration in abundance of U4, U5, and U6, observed in our study, contributes to the understanding of the fact that, the metabolism of major spliceosomal UsnRNAs is differentially regulated during the process of neoplastic transformation.
In the present study, we have experimentally and theoretically studied the free-radical quenching... more In the present study, we have experimentally and theoretically studied the free-radical quenching property of dihydrocanaric acid (DCA) isolated from seedpods of Holarrhena antidysenterica. A modified method was used to estimate the nitric oxide scavenging effect of the DCA (significant activity of 75.22%) along with methanolic extract of seed pods of Holarrhena antidysenterica (72.80%) compared to the ascorbic acid as standard (40.60%). Studies have also been conducted for superoxide scavenging activity of the DCA (78.82%) and methanolic extract of seed pods (84.28%) compared to quercetin as standard (82.08%). Theoretically, it has been determined by density-functional theory(DFT) calculations using M06-2X hybrid functional and the double-ζ- split-valence 6-31G (d, p) basis set that the nitric oxide scavenging activity of the compound is by the addition of NO radical at double bond position. Predicted biological activity profile of DCA suggests that it has less activity probability (Pa) for toxicity (Pa = 0.730), cytotoxicity (Pa = 0.208), compared to those chemical entities that are already known as anticancer agents indicating that DCA is less toxic and more tolerable for normal cells. Furthermore, molecular docking studies of the DCA with different studied cancer-related receptors [Estrogen receptor (− 60.12 kcal/mol), epidermal growth factor receptor (EGFR) (− 30.33 kcal/mol), estrogen receptor alpha (− 4.82 kcal/mol), uPAR (− 32.55 kcal/mol) and an enzyme having lipid kinase activity phosphoinositide 3-kinase (− 55.94 kcal/mol)] were found to have better binding affinities compared to betulinic acid and doxorubicin. Thus, our findings suggest that the DCA could be a safer and effective alternative in fighting cancer with minimal side effects.Electronic supplementary materialThe online version of this article (10.1007/s13205-020-02221-5) contains supplementary material, which is available to authorized users.
Atlas of genetics and cytogenetics in oncology and haematology, Aug 1, 2017
B-cell leukemia/lymphoma 1) 130 Leukaemia Section Classification of myelodysplastic syndromes 201... more B-cell leukemia/lymphoma 1) 130 Leukaemia Section Classification of myelodysplastic syndromes 2015 155 Virginie Eclache t(11;19)(q13;p13) FSTL3/CCND1 162 Jean-Loup Huret Case Report Section The t(2;12)(p12;p13), leading to the juxtaposition of the cyclin D2 CCND2 gene with the immunoglobulin kappa locus (IGK), defines a small subset of mantle cell lymphomas lacking cyclin D1 expression.
Cell culture and animal studies have demonstrated strong chemopreventative effects of green tea a... more Cell culture and animal studies have demonstrated strong chemopreventative effects of green tea and its associated polyphenols in multiple cancers, though the exact mechanisms of action are not well understood. This in vitro study examined the antiproliferative/pro-apoptotic potential of green tea extract (GTE), polyphenon-60 (PP-60), (−)-epicatechin gallate (ECG) and (−)-epigallocatechin-3-gallate (EGCG) in both normal and malignant human bladder cells. Cell growth (proliferation/ apoptosis) was measured in UROtsa (normal), SW780 (tumorigenic; low-grade), and TCCSUP (tumorigenic; high-grade) human bladder urothelial cells by cell proliferation (XTT) assay after treatment with 0-80 μg/mL of GTE, PP-60, ECG and EGCG for 72 h. Molecular signaling pathways of catechin-induced apoptosis were analyzed using Human signal transduction RT 2 Profiler PCR array (SuperArray). Compared to control-treated cells, treatment with catechin agents significantly suppressed cell growth in a dose-dependent fashion (P < 0.01), with strongest effects evoked by ECG and EGCG in UROtsa cells, ECG in low-grade RT4 and SW780 cells, and PP-60 and EGCG in high-grade TCCSUP and T24 cells. Microarray analysis indicated distinct differences in mRNA gene expression regarding growth signaling pathway activation induced by EGCG in normal/tumorigenic human bladder cell lines, providing a rationale for the putative therapeutic usage of green tea polyphenols against bladder disease.
Head and neck squamous cell carcinoma (HNSCC) is the most common and fatal cancer type in India w... more Head and neck squamous cell carcinoma (HNSCC) is the most common and fatal cancer type in India with about 30–40% of frequency. Molecular etiologies of the disease include consumption of alcohol, tobacco habit and HPV infection. The lack of understanding in molecular pathogenesis of the disease leads to poor survival of HNSCC patients. Cytogenetic analysis has shown that the chromosomal abnormalities gradually increase with progression of the tumor in a non-random manner. Molecular analysis of- copy number variation, promoter methylation, mutation and expression (gene and protein) revealed identification of some of the candidate tumor suppressor genes located in the chromosomal 3p, 8q, 9p/q, 11q, 13q and 17p etc. regions associated with development of HNSCC. These genes are further seen to control different cellular pathways like- (1) frequent inactivation of SH3GL2 and CDC25A- associated with EGFR homeostasis and de-phosphorylation respectively; (2) Rb and p53 associated pathways in G1-S cell cycle transition; (3) Slit—ROBO associated signaling pathways; (4) PTCH associated Hedgehog signaling; (5) DNA repair pathways like- mismatch and double strand break repair; (6) apoptosis and (7) integrin signaling. Now, systemic approach has been taken to characterize the molecular pathways in this tumor to find out the key regulatory steps for management of the disease.
Breast cancer (BC) is frequent among women in worldwide as well as in India. Several studies have... more Breast cancer (BC) is frequent among women in worldwide as well as in India. Several studies have reported a wide variation (1.6-86.2%) in the frequency of incidence of human papillomavirus (HPV) infection in BC with high prevalence of high risk HPV16 subtype. HPV infection in breast can occur through different routes like body fluid or by micro-lesion of breast skin from genital/agential sites, though the actual mode of HPV transmission is not yet known in details. Frequent integration and sequence variation with low copy number of HPV16 were seen in this tumour. In addition, high frequencies of methylation in p97 promoter region of HPV16 were evident in this tumour. Novel splice variants of E6/E7 along with other common variants and their protein expression were seen in the tumour. This indicates the importance of HPV in this tumor, its early diagnosis and prognosis. Thus, HPV may be targeted through vaccination to control the disease. However, detailed analysis of HPV associated molecular pathogenesis of BC is warranted for proper therapeutic intervention.
Indian Journal of Medical and Paediatric Oncology, Apr 1, 2021
Introduction The improved prognosis of pediatric B-cell acute lymphoblastic leukemia (pBALL) is c... more Introduction The improved prognosis of pediatric B-cell acute lymphoblastic leukemia (pBALL) is considered as a good progress of medical science in the field of oncology and hematology. Minimal residual disease (MRD) refers to presence of disease in molecular level is a newer practice with respect to the detection of complete remission by conventional pathologic analysis. Prognostic value of MRD in pediatric ALL (p-ALL) is well known. Objectives This study was aimed to describe clinical outcomes and prognosis, that is, overall survival and relapse in the patients with pBALL with respect to minimal residual disease detection on day 15, day 29, and postconsolidations in a tertiary care center in eastern India. Materials and Methods Eight color flow cytometry was used to detect MRD in this study. This contained markers such as CD 19, CD 34, CD 10, CD58, CD 45, CD13, anti-TDT, CD33. Eight panels included were (1) CMPO-FITC/cCD79a-PE/cCd3ECD, (2) CD20-FITC/cCD10-PE/cCd-19ECD, (3) CD34-FITC/cCD117-PE/cCd45 ECD/CD2 PC 5, (4) CD15 FITC/ CD33PE/CD45ECD, (5) CD14 FITC/CD13 PE/CD45ECD, (6) HLADR FITC/ CD7 PE/CD45 ECD, (7) TdT FITC/CD45 ECD (IF CD34 NEG), and (8) CD58 FITC/CD 45 ECD (IF BOTH CD34 AND TdT NEG; were used to prepare the marker. Results The study included 52 patients. In the 52 patients, 59.6% patients are alive with a p-value of 0.031. MRD was checked on every 15th and the 29th day and postconsolidation of the treatment where in day 15 (p = 0.023), it was 53.4% positive and 46.5% negative. On day 29 (p = 0.031), MRD was 22.5% positive and 77.5% negative, in post consolidation, it was positive in 20% and negative is 80%. MRD value below 0.01 is taken as negative and above is taken as positive. The overall survival (OS) is of 32.88 + 8.59 with a 6 to 36 months of duration. In relapsed cases, no hemorrhagic relapse was found and two CNS relapses were found. Conclusion It was a study of 52 patients of pBALL with a detection of MRD by FCM. MRD-negative patients had a good prognosis and comparatively lower rate of relapse than the one with positive MRD. Effort should be made to adhere to recommendation of MRD testing in clinical guidelines.
Mouse plasmacytomas (MPCs) induced by pristane oil, or by a combination of pristane oil and Abels... more Mouse plasmacytomas (MPCs) induced by pristane oil, or by a combination of pristane oil and Abelson virus, carry one of two chromosomal translocations. The typical 12; 15 translocation leads to the juxtaposition of c-myc and immunoglobulin heavy-chain sequences, whereas the 6; 15 translocation links the kappa light-chain locus with the pvt-1 (plasmacytoma variant translocation) locus, located at least 75kb 3' of c-myc [Cory, S., Graham, M., Webb, E., Corcoran, L. & Adams, J. (1985). EMBO J., 4, 675-681]. Unlike the human Burkitt's lymphoma-associated translocation, the lambda/myc juxtaposed variant translocation has not been found previously in MPCs. Using unconventional MPC induction systems in which the tumor precursor cell was induced to proliferate in a secondary host, we have recently identified a 15; 16 translocation in six of the derived MPCs [Wiener, F., Silva, S., Sugiyama, H., Babonits, M. & Klein, G. (1990). Genes Chromosomes Cancer, 2, 36-43]. Chromosome 16 harbors the lambda light-chain gene. To explore whether the 15; 16 translocation represents the lambda/myc juxtaposition, we have mapped the breakpoints on chromosomes 15 and 16 by pulsed-field gel electrophoresis (PFGE). The pvt-1 region was mapped to approximately 220 kb 3' of c-myc. The breakpoint on chromosome 15 in ABPC-Ch-163-10, one of the six 15; 16 translocation-carrying MPCs, was situated approximately 80 kb 3' of c-myc and 140 kb 5' of pvt-1b, the major breakpoint cluster region of the previously analysed 6; 15 variant MPCs. The breakpoint on chromosome 16 was found to cut between the V1 and C3 regions of the lambda locus. Co-migration experiments showed that the C3 and the myc gene were juxtaposed head to tail on the 15; 16 translocation chromosome. On the reciprocal product V1 was juxtaposed to pvt-1.
Background & objectives: Medicinal plants like Swertia chirata are rich sources of different ... more Background & objectives: Medicinal plants like Swertia chirata are rich sources of different xanthones. This study was aimed to assess the cytotoxic potential of four most abundant xanthones present in S. chirata both in vivo and in vitro in Ehrlich ascites carcinoma (EAC), a mouse transplantable breast carcinoma cell line and two human breast carcinoma cell lines (MCF-7 and MDA-MB-231). Methods: Four xanthones derived from S. chirata namely 1-hydroxy-3,7,8-trimethoxyxanthone (XA), 1,8-dihydroxy-3,5-dimethoxyxanthone (XB), 1-hydroxy-3,5,8-trimethoxyxanthone (XC) and 1,5,8-trihydroxy-3-methoxyxanthone (XD) were used for determination of sub-lethal dose on the cell lines EAC, MCF-7, MDA-MB-231 and verified toxicity of sub-lethal dose on normal murine fibroblast cells. Cytotoxicity was measured in vivo and survivability of mice was plotted accordingly. Therapeutic efficacy of XD was evaluated both in vivo and in vitro by determination of lipid peroxidation (LPO), reactive oxygen species (ROS) generation and by quantitating the enzyme status (GSH, catalase, superoxide dismutase) in treated and untreated samples. DNA damage was evaluated using comet and DNA fragmentation assays. Furthermore, apoptotic effect was analyzed by flow cytometry and validated by TUNEL assay and Western blotting. Results: Among all the xanthones tested XD showed IC50at the lowest dose, and normal cells were unaffected at this dose. Survivability of mice increased significantly when treated with XD compared to other xanthones and cisplatin. Significantly increased ROS and LPO were found in cancer cells as a result of XD treatment which was unaltered in normal cell line. XD induced DNA damage and apoptosis in cancer cell lines. Interpretation & conclusions: Our experimental data indicate that XD may potentially act as a chemotherapeutic agent by enhancing ROS in breast cancer cells thereby leading to apoptosis.
The present study was designed to determine the chemopreventive efficiency of 1,5,8-trihydroxy-3-... more The present study was designed to determine the chemopreventive efficiency of 1,5,8-trihydroxy-3-methoxy xanthone, abbreviated as TMX, isolated and purified from the aerial part of the plant Swertia chirata against 9,10-dimethylbenz[a]-anthracene (DMBA)/croton oilinduced skin cancer, and probe into the molecular mechanism. All the mice in the carcinogen control group developed severe dysplastic lesions after the 14th week of application of the carcinogen, which progressed to carcinoma in situ around the 20th week; this was validated histologically. However, after TMX treatment, only around 50% of mice developed papilloma which histologically was found to be restrictive to moderate to severe hyperplastic change in the 14th and 20th week. The chemopreventive potential was determined by calculating the attributable risk (AR), which was-11.3 for the 14th week and increased up to-17.5 for the 20th week. To ascertain the effect of TMX treatment on inflammation, the effect of TMX on inflammatory cytokines was studied by ELISA. It revealed a significant reduction in inflammation upon TMX treatment for the 20th week. As TMX could hold its chemopreventive potential up to the 20th week, the molecular mechanism of restriction was studied for the 20th week of treatment. Skin forms a rich source of stem cells which orchestrate the progression of carcinogenesis and become cancer stem cells (CSCs). β-Catenin and KRAS are known central modulators of CSCs, that play a crucial role in the progression of skin carcinogenesis. We observed that TMX treatment inhibited KRAS and nuclear translocation of β-catenin causing its cytoplasmic degradation by P-53 and P-21mediated pathway, thereby exerting its chemopreventive potential.
Head and neck squamous cell carcinoma is one of the leading cancers in terms of incidence and mor... more Head and neck squamous cell carcinoma is one of the leading cancers in terms of incidence and mortality. However, no reliable marker till date accurately predicts its progression when altered in healthy tissues. The study aims to identify alleles of microsatellites adjacent to important cell cycle regulatory, tumor suppressor genes altered in early head and neck lesions, viz. RBSP3, LIMD1 and CDC25A, which undergo frequent deletion and can be used for population screening and early detection. DNA for tumors and normal tissues was isolated from 143 patients in different stages of head and neck squamous cell carcinoma. The size of microsatellite present in normal tissues and their deletion in the corresponding tumor was identified, along with the correlation of expression in normal epithelium with respect to allele size. The results revealed a range of alleles (CA 9 to CA 32) for the different microsatellites of the genes in normal tissues. The larger alleles were significantly deleted with differential deletion of alleles observed in tumors, except for LIMD1, in which the smaller allele was significantly deleted. In normal tissues, some alleles represented as stable alleles with high prevalence, while in tumours, specific sizes showed greater propensity for deletion. However, similar expression of the proteins in normal epithelium adjacent to tumors was observed despite variations in allele size, possibly due to the location of the microsatellites. Thus, those alleles when present in normal tissues and undergoing persistent deletion in tumours could be used as markers for screening and early identification of populations at risk of developing head and neck lesions.
Background and Objectives: The aim of the study was to understand the importance of mismatch repa... more Background and Objectives: The aim of the study was to understand the importance of mismatch repair genes MLH1 and MSH2 in chemotolerance and prognosis of breast carcinoma (BC). Methods: First, the alterations (deletion/methylation/expression) of MLH1 and MSH2 were analyzed in 45 neoadjuvant chemotherapy (NACT)-treated and 133 pretherapeutic BC samples. The chemotolerant BC cells were characterized by treating two BC cell lines MCF-7 and MDA MB 231 with two anthracycline antitumor antibiotics, doxorubicin and nogalamycin. Results: The deletion frequencies were 32% to 38% in MLH1/MSH2 genes and promoter methylation frequencies were 49% to 62% in MLH1 and 41% to 51% in MSH2 in both NACT-treated and pretherapeutic samples. The overall alteration of MLH1 and MSH2 was 58% to 71% in the samples. Reduced messenger RNA (mRNA) and protein expression were found in both the genes and it showed concordance with the molecular alterations. NACT-treated patients showed better prognosis. The chemotherapeutic drug induced increased mRNA/protein expression of the genes in BC cell lines was due to their promoter hypomethylation, as analyzed by quantitative methylation assay. This phenomenon was also evident in NACT-treated BC samples. Conclusion: MLH1/MSH2 genes play a critical role in the development of BC. Hypomethylation of MLH1/MSH2 genes might be important in chemotolerance of the disease.
Background: Genetic instability of chromosome 11 is a frequent event in many solid tumours, inclu... more Background: Genetic instability of chromosome 11 is a frequent event in many solid tumours, including head and neck squamous cell carcinoma (HNSCC). Aims: To perform allelic imbalance analysis of cytogenetically mapped altered regions of human chromosome 11 in patients with HNSCC from eastern India. Methods: Genomic alterations were investigated using highly polymorphic microsatellite markers in both HNSCC and leukoplakia tissues. Results: Microsatellite markers D11S1758 from 11p13-15 and D11S925 from 11q23.3-24 had the highest frequency (38% and 32%, respectively) of loss of heterozygosity among all the markers analysed. Allelic loss at the marker D11S925 was seen in both leukoplakia and in all stages of HNSCC tumour tissues suggesting that it is an early event in HNSCC tumorigenesis. Microsatellite size alteration was also found to be high (> 20%) in several markers. In leukoplakia samples microsatellite instability was seen at a higher frequency than loss of the allele, indicating such alterations might initiate the process of tumorigenesis in HNSCC. Conclusions: The high rate of chromosomal alterations at 11q21-24 in HNSCC suggests the presence of a putative tumour suppressor gene in this region.
Purpose In approximately 30% of triple-negative breast cancer (TNBC) patients a complete patholog... more Purpose In approximately 30% of triple-negative breast cancer (TNBC) patients a complete pathological response is achieved. However, after neo-adjuvant chemotherapy treatment (NACT) residual tumour cells can be intrinsically resistant to chemotherapy. In this study, associations of the WNT/beta-catenin pathway with chemo-tolerance of NACT treated TNBC patients were compared to that of pre-treatment TNBC patients. Methods Expression analyses were performed in both pre-treatment and NACT treated TNBC samples using immunohistochemistry and qRT-PCR, along with DNA copy number variation (CNV) and promoter methylation analyses to elucidate the mechanism(s) underlying chemo-tolerance. In addition, in vitro validation experiments were performed in TNBC cells followed by in vivo clinicopathological correlation analyses. Results A reduced expression (41.1%) of nuclear beta-catenin together with a low proliferation index was observed in NACT samples, whereas a high expression (59.0%) was observed in pre-treatment samples. The reduced nuclear expression of beta-catenin in the NACT samples showed concordance with reduced expression levels (47-52.9%) of its associated receptors (FZD7 and LRP6) and increased expression levels (35.2-41.1%) of its antagonists (SFRP1, SFRP2, DKK1) compared to those in the pre-treatment samples. The expression levels of the receptors showed no concordance with its respective gene copy number/mRNA expression statuses, regardless treatment. Interestingly, however, significant increases in promoter hypomethylation of the antagonists were observed in the NACT samples compared to the pre-treatment samples. Similar expression patterns of the antagonists, receptors and beta-catenin were observed in the TNBC-derived cell line MDA-MB-231 using the anthracyclines doxorubicin and nogalamycin, suggesting the importance of promoter hypomethylation in chemotolerance. NACT patients showing reduced receptor and/or beta-catenin expression levels and high antagonist expression levels exhibited a comparatively better prognosis than the pre-treatment patients. Conclusions Our data suggest that reduced nuclear expression of beta-catenin in NACT TNBC samples, due to downregulation of its receptors and upregulation of its antagonists through promoter hypomethylation of the WNT pathway, plays an important role in chemo-tolerance.
Atlas of genetics and cytogenetics in oncology and haematology, Nov 1, 2011
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 ... more This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence.
Differential alterations of the spliceosomal Uridylic acid rich small nuclear RNAs (UsnRNAs) (U1,... more Differential alterations of the spliceosomal Uridylic acid rich small nuclear RNAs (UsnRNAs) (U1, U2, U4, U5, and U6) are reported to be associated with cellular proliferation and development, but definitive information is scarce and also elusive. An attempt is made in this study to analyze the metabolic patterns of major spliceosomal UsnRNAs, during tumor development, in an in vitro carcinogenesis model of 20-methylcholanthrene (MCA)-transformed Swiss Mouse Embryonic Fibroblast (MEF), designated as CNCI-PM-20. MEF cells, after treatment with 20-MCA, progressed through a sequence of passages with distinct and heritable changes, finally becoming neoplastic at passage-42 (P42). A differential expression pattern of major UsnRNAs was observed during this process. The abundance of U1 was 20% below control (P1) at passage-20 (P20), followed by a gradual increase up until P42 ($12% above the P1 value). The abundance of U2 was more or less constant during the cellular transformation. U4 showed a trend of increase, with above 30% abundance than control at P20, followed by a significant increase at P36 and P42 (1.5-and 2-fold, respectively, P-value <0.01). U5 also followed an identical pattern, with an increase of 70% compared to control (P-value <0.05) at P42. Interestingly, U6 gradually decreased from P20 onwards up until P42, with 22% at P20 and 67% at P42 (P-value <0.01). An overall significant quantitative alteration in abundance of U4, U5, and U6, observed in our study, contributes to the understanding of the fact that, the metabolism of major spliceosomal UsnRNAs is differentially regulated during the process of neoplastic transformation.
In the present study, we have experimentally and theoretically studied the free-radical quenching... more In the present study, we have experimentally and theoretically studied the free-radical quenching property of dihydrocanaric acid (DCA) isolated from seedpods of Holarrhena antidysenterica. A modified method was used to estimate the nitric oxide scavenging effect of the DCA (significant activity of 75.22%) along with methanolic extract of seed pods of Holarrhena antidysenterica (72.80%) compared to the ascorbic acid as standard (40.60%). Studies have also been conducted for superoxide scavenging activity of the DCA (78.82%) and methanolic extract of seed pods (84.28%) compared to quercetin as standard (82.08%). Theoretically, it has been determined by density-functional theory(DFT) calculations using M06-2X hybrid functional and the double-ζ- split-valence 6-31G (d, p) basis set that the nitric oxide scavenging activity of the compound is by the addition of NO radical at double bond position. Predicted biological activity profile of DCA suggests that it has less activity probability (Pa) for toxicity (Pa = 0.730), cytotoxicity (Pa = 0.208), compared to those chemical entities that are already known as anticancer agents indicating that DCA is less toxic and more tolerable for normal cells. Furthermore, molecular docking studies of the DCA with different studied cancer-related receptors [Estrogen receptor (− 60.12 kcal/mol), epidermal growth factor receptor (EGFR) (− 30.33 kcal/mol), estrogen receptor alpha (− 4.82 kcal/mol), uPAR (− 32.55 kcal/mol) and an enzyme having lipid kinase activity phosphoinositide 3-kinase (− 55.94 kcal/mol)] were found to have better binding affinities compared to betulinic acid and doxorubicin. Thus, our findings suggest that the DCA could be a safer and effective alternative in fighting cancer with minimal side effects.Electronic supplementary materialThe online version of this article (10.1007/s13205-020-02221-5) contains supplementary material, which is available to authorized users.
Atlas of genetics and cytogenetics in oncology and haematology, Aug 1, 2017
B-cell leukemia/lymphoma 1) 130 Leukaemia Section Classification of myelodysplastic syndromes 201... more B-cell leukemia/lymphoma 1) 130 Leukaemia Section Classification of myelodysplastic syndromes 2015 155 Virginie Eclache t(11;19)(q13;p13) FSTL3/CCND1 162 Jean-Loup Huret Case Report Section The t(2;12)(p12;p13), leading to the juxtaposition of the cyclin D2 CCND2 gene with the immunoglobulin kappa locus (IGK), defines a small subset of mantle cell lymphomas lacking cyclin D1 expression.
Cell culture and animal studies have demonstrated strong chemopreventative effects of green tea a... more Cell culture and animal studies have demonstrated strong chemopreventative effects of green tea and its associated polyphenols in multiple cancers, though the exact mechanisms of action are not well understood. This in vitro study examined the antiproliferative/pro-apoptotic potential of green tea extract (GTE), polyphenon-60 (PP-60), (−)-epicatechin gallate (ECG) and (−)-epigallocatechin-3-gallate (EGCG) in both normal and malignant human bladder cells. Cell growth (proliferation/ apoptosis) was measured in UROtsa (normal), SW780 (tumorigenic; low-grade), and TCCSUP (tumorigenic; high-grade) human bladder urothelial cells by cell proliferation (XTT) assay after treatment with 0-80 μg/mL of GTE, PP-60, ECG and EGCG for 72 h. Molecular signaling pathways of catechin-induced apoptosis were analyzed using Human signal transduction RT 2 Profiler PCR array (SuperArray). Compared to control-treated cells, treatment with catechin agents significantly suppressed cell growth in a dose-dependent fashion (P < 0.01), with strongest effects evoked by ECG and EGCG in UROtsa cells, ECG in low-grade RT4 and SW780 cells, and PP-60 and EGCG in high-grade TCCSUP and T24 cells. Microarray analysis indicated distinct differences in mRNA gene expression regarding growth signaling pathway activation induced by EGCG in normal/tumorigenic human bladder cell lines, providing a rationale for the putative therapeutic usage of green tea polyphenols against bladder disease.
Head and neck squamous cell carcinoma (HNSCC) is the most common and fatal cancer type in India w... more Head and neck squamous cell carcinoma (HNSCC) is the most common and fatal cancer type in India with about 30–40% of frequency. Molecular etiologies of the disease include consumption of alcohol, tobacco habit and HPV infection. The lack of understanding in molecular pathogenesis of the disease leads to poor survival of HNSCC patients. Cytogenetic analysis has shown that the chromosomal abnormalities gradually increase with progression of the tumor in a non-random manner. Molecular analysis of- copy number variation, promoter methylation, mutation and expression (gene and protein) revealed identification of some of the candidate tumor suppressor genes located in the chromosomal 3p, 8q, 9p/q, 11q, 13q and 17p etc. regions associated with development of HNSCC. These genes are further seen to control different cellular pathways like- (1) frequent inactivation of SH3GL2 and CDC25A- associated with EGFR homeostasis and de-phosphorylation respectively; (2) Rb and p53 associated pathways in G1-S cell cycle transition; (3) Slit—ROBO associated signaling pathways; (4) PTCH associated Hedgehog signaling; (5) DNA repair pathways like- mismatch and double strand break repair; (6) apoptosis and (7) integrin signaling. Now, systemic approach has been taken to characterize the molecular pathways in this tumor to find out the key regulatory steps for management of the disease.
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