Inhibition of NOTCH signaling pathway chemosensitizes HCC CD133+ cells to vincristine and 5-fluorouracil through upregulation of BBC3.

In hepatocellular carcinoma (HCC), the poor response to the chemotherapeutic agents is partially attributed to the chemoresistance property of cancer stem cells (CSCs). NOTCH signaling pathway plays a crucial role in the chemoresistance through the maintenance of the CSCs. We observed that the NOTCH pathway was activated in HCC CD133+ cells treated with vincristine (VIN)1 and 5-fluorouracil (5-FU)2. Therefore, we examined whether inhibition of the NOTCH can improve sensitization of HCC CD133+ cells to VIN and 5-FU. The Huh7 cell line was pre-incubated γ-secretase DAPT, as a NOTCH inhibitor, and then treated with IC50 dose of VIN or 5-FU. The CD133+ cells were then isolated and analyzed for the cell viability, apoptosis, migration and spheroid formation capacities, and gene and protein expression. It was observed that pre-incubation with DAPT significantly downregulated the expression of NOTCH-related genes and led to a significant reduction in VIN- and 5-FU-CD133+ population. In addition, DAPT pre-incubated VIN- and 5-FU-treated-CD133+ cells formed fewer spheroids in 3D culture and had a lesser migration capacity in 2D culture. Importantly, DAPT enhanced the apoptosis rate of VIN- and 5-FU-treated CD133+ cells for 3- and 2-fold, which was correlated with the enhanced expression of pro-apoptotic BBC3 (BCL-2-binding component 3) and decreased expression of HES1 that was reported to regulate BBC3 negatively. Collectively, it was observed that NOTCH inhibition sensitized the HCC CD133+ cells to VIN and 5-FU through enhancing BBC3-mediated apoptosis. The results highlighted the role of NOTCH/HES1/BBC3 axis in resistance of CD133+ cells to VIN and 5-FU. Understanding the molecular mechanisms underlying chemoresistance in HCC CD133+ cells may help in designing the novel targeted therapies to chemosensitize them.

Impact of single nucleotide polymorphism in chemical metabolizing genes and exposure to wood smoke on risk of cervical cancer in North-Indian women.

AIM: In the present study, we investigated the hypothesis whether exposure to wood smoke increases the risk of cervical cancer (CC) in North-Indian women who inherit different polymorphic forms of chemical metabolizing genes (GSTM1, GSTT1, GSTP1 and CYP1A1). MATERIALS AND METHODS: One hundred fifty histologically confirmed CC patients and equal number of cancer-free age and ethnicity matched controls were genotyped for genetic polymorphism in chemical metabolizing genes by using polymerase chain reaction/restriction fragment length polymorphism method. The association of the different genotypes and exposure to wood smoke with the risk of CC in North-Indian women was estimated by doing statistical analysis using Statistical Package for the Social Science. RESULTS: It was observed that the variant genotypes of GSTM1, GSTT1, GSTP1 and CYP1A1 did not significantly increase the risk of CC. However, statistically significant increased risk (odds ratio 3.6; 95% confidence interval, 1.34-9.78; p = 0.008) was observed for women who used wood for cooking and had GSTM1 (null) genotype. CONCLUSIONS: The present study suggests that genetic differences in the metabolism of wood smoke carcinogens, particularly by GSTM1, may increase the risk of CC.

The protective role of the -1306C>T functional polymorphism in matrix metalloproteinase-2 gene is associated with cervical cancer: implication of human papillomavirus infection.

Cervical cancer is the major reproductive health problem among women caused by persistent infection of high-risk human papillomavirus (HR-HPV). Metalloproteinase-2 (MMP-2) is an endopeptidase highly expressed in cervical cancer; however, the genetic link between aberrant expression of MMP-2 and cervical carcinogenesis is not known. The genotypic distribution, expression pattern of MMP-2 and HPV infection, was analyzed in a total of 300 fresh surgically resected cervical tissue biopsies. The MMP-2 C1306T (rs243865) promoter polymorphism dominant model (CC v/s CT + CT + TT) revealed that the CC genotype had a 4.33-fold significant increased risk for development of cervical cancer (OR = 4.33; 95 % CI = 2.36-4.02, p = 0.0001) compared to those with variant genotypes (-1306 CT + TT). The C allele was associated with 3-fold significant increased risk (OR = 2.95; 95 % CI = 1.90-4.60, p = 0.0002) compared to T allele. Interestingly, a significant correlation was found between high expression of MMP-2 protein and CC genotype in cancer patients (p = 0.001) compared to normal controls (p = 0.012). Further analysis showed that the risk of cancer was extremely pronounced in HPV positive patients (OR = 9.33; 95 % CI = 2.88-30.20, p = 0.0001) compared to HPV negative ones, implicating the possible interaction between -1306CC genotype and HPV infection in increasing the cancer risk (p = 0.0001). The leads from the present study suggest the protective role of gene variant -1306C>T at the promoter region of the MMP-2 against HPV-mediated cervical cancer. These findings substantiate the functional role of MMP-2 C1306T polymorphism in a significant downregulation of MMP-2 protein in women with variant genotype (CT/TT) compared to the normal wild CC genotype.

Analysis of genetic variations across regulatory and coding regions of kappa-casein gene of Indian native cattle (Bos indicus) and buffalo (Bubalus bubalis).

The promoter region of kappa-casein (κ-CN) gene in Indian native cattle and buffalo breeds was sequenced and analyzed for nucleotide variations. Sequence comparison across breeds of Indian cattle revealed a total of 7 variations in the promoter region, of which - 515 G/T, - 427 C/T, - 385 C/T, - 283 A/G and - 251 C/T were located within consensus binding sites for octamer-binding protein (OCT1)/pregnancy specific mammary nuclear factor (PMF), activator protein-2 (AP2), hepatocyte nuclear factor (HNF-1) and GAL4 transcription factors (TFs), respectively. These variations might be involved in gain or loss of potential transcription factor binding sites (TFBSs). Unlike the other 4 variants, the - 283 (A/G) variant located within HNF-1 TFBS was specific to Indian cattle as this change has not been observed in the Bos taurus sequence. Other TFBSs viz., MGF, TBP, NF-1, milk box and C/EBP were conserved across species. For the Indian native buffalo breeds, only 3 changes were identified in the promoter region; - 305 (A/C), - 160 (T/C) and - 141 (A/G) and most of the TFBSs were found to be conserved. However, deletion of two adjacent nucleotides located in and around binding site for C/EBP TF was identified in buffalo when compared with promoter sequence of bovine κ-CN. For κ-CN of Indian native cattle, a strong linkage disequilibrium (LD) was observed for variations 515 G/T, - 427 C/T and - 385 C/T in the promoter region; and for variations at codons 136 and 148 of exon-IV. Further, among intragenic haplotypes, variation - 427 C/T was found to be in LD with variations at codons 136 and 148. The information generated in the present work provides comprehensive characterization of κ-CN gene promoter and coding regions in Indian cattle and buffaloes and reported variations could become important candidates for carrying out further research in dairy traits.

Sequence analysis and identification of new variations in the 5'-flanking region of αS2-casein gene in Indian zebu cattle.

Regulatory region of milk protein alpha S2-casein (αS2-CN) gene sequence was characterized and analyzed for nucleotide variations in animals representing 13 Indian zebu cattle (Bos indicus) breeds. A total of 15 variations; 11 in promoter region (1.56 Kb): -1481 (C>T), -1412 (C>T), -1342 (C>T), -1084 (G>A), -979 (A>G), -657 (A>T), -508 (A>G), -186 (T>C), -184 (T>C), -151 (T>C) and -135 (C>T); 1 in 5'-UTR (44 bp): 7 (C>T) while, 3 in intron-I region (73 bp): 186 (C>T), 194 (A>C) and 301 (A>T) were identified. Additionally, single deletion was observed at -975 (A>-) but not involve any known potential transcription factor binding sites (TFBS). Comparison with Bos taurus sequence revealed two additional variations -1085 (T>C) and -739 (A>G). Out of the total 18 variations observed between indicine and taurine αS2-CN regulatory region sequence, 15 were novel to B. indicus and are reported for the first time. Among these, four variations were located within the potential TFBSs; -1342 (C>T) within HNF-3beta, -739 (A>G) within C/EBP-alpha while -657 (A>T) and -508 (A>G) were found within glucocorticoid receptor TFBSs. Variations located within or in proximity to putative TFBSs could possibly influence the binding affinity of nuclear factors towards DNA binding domains, thus affecting transcriptional rate of αS2-CN gene. Phylogenetically, as expected, Indian zebu cattle were grouped close to B. taurus and were most distantly placed in comparison to human. The study indicated possible genetic variations in the regulatory regions of αS2-CN gene within Indian native cattle (B. indicus) and also its comparison with evolutionary different B. taurus breeds.

Downregulation of tumor suppressor gene PML in uterine cervical carcinogenesis: impact of human papillomavirus infection (HPV).

OBJECTIVES: Cervical cancer is a leading gynecological cancer in Indian women and is caused due to infection with high risk human pappilloma virus (HR-HPV) 16 and 18. It has been well documented that PML (promyelocytic leukemia) enhances viral infectivity and plays a crucial role in antiviral response mechanisms. The aim of the present study was to evaluate the role of PML gene with context to HPV infection in cervical carcinogenesis. METHODS: The expression pattern of PML was analyzed by western blotting and immunohistochemistry in a total of 170 fresh surgically resected cervical tissue specimens comprising precancer (n=12), cancer (n=118) and normal controls (n=40) recruited from PGIMER, Chandigarh, India. HPV status was analyzed by L1 consensus PCR followed by type specific PCR for HR-HPV types 16 and 18 and low risk types 6 and 11. RESULTS: A significant downregulation of PML protein was observed in the majority of cervical cancer and precancer cases 68% (89/130) compared to normal controls. The loss of expression pattern of PML gene was significantly increased with severity of disease both clinically and pathologically (p<0.001). HPV infection was detected in the majority of cancer cases 96% (113/118) and in 83% (10/12) of precancer lesions whereas no infection could be detected in normal controls. Interestingly, all the 68% (89/130) cervical cancer cases that showed downregulation of PML were HPV infected (p=0.0001). CONCLUSION: Taken together, these observations suggest that the downregulation of PML gene and its synergism with HPV infection may play an important role and may serve as a new marker for early diagnosis and therapeutic intervention for cervical carcinogenesis.

The -137G/C polymorphism of interleukin 18 promoter and risk of HIV-1 infection and its progression to AIDS.

A growing body of evidence suggests that host genetic factors play an important role both in susceptibility to human immunodeficiency virus 1 (HIV-1) infection and in progression to AIDS. Interleukin 18 (IL-18) is a pleiotropic proinflammatory cytokine that serves as an important regulator of immune responses. It plays a key role in induction of both Th1 and Th2 cytokines and, thereby, modulates their immune responses. Single nucleotide polymorphisms in the IL-18 gene promoter region may lead to an altered transcriptional activity and IL-18 production, and so this may account for individuals' variation to the risk of HIV-1 infection. With this perspective, the -137G/C polymorphism in the promoter region of the IL-18 gene was studied in 500 patients with HIV-1/AIDS and an equal number of sex and age matched healthy controls using sequence specific polymerase chain reaction analysis. We did not observe any significant association of the heterozygous G/C genotype with the risk of HIV-1-infection/AIDS. However, statistically significant associations of the G allele and homozygous G/G genotype of -137 G/C polymorphism of IL-18 promoter with increased risk of HIV-1/AIDS were identified. The data of the present study suggest that IL-18 -137 G allele and G/G genotype seem to be involved in the pathogenesis of HIV-1 infection among North Indians.

Aberrant promoter methylation and loss of suppressor of cytokine signalling-1 gene expression in the development of uterine cervical carcinogenesis.

BACKGROUND: Cervical cancer is a leading cause of cancer related deaths in women worldwide caused due to infection of high-risk human papillomaviruses. As JAK/STAT signalling pathway has been shown to play an important role during carcinogenesis, we studied the role of silencing of Suppressor of Cytokine Signalling-1 (SOCS-1) gene, a negative regulator of JAK/STAT pathway in cervical cancer. METHODS: The expression pattern of SOCS-1 mRNA and protein was analyzed in different stages of cervical tumor biopsies while normal cervical tissues served as controls. RT-PCR, immunohistochemistry and methylation-specific PCR (MSP) were performed to assess the expression pattern and promoter methylation of SOCS-1 gene in a total of 120 fresh surgically resected cervical tissue specimens comprising precancer (n = 12), cancer (n = 78) and normal controls (n = 30). RESULTS: Compared with expression of SOCS-1 in normal tissues, 64% of the tumor tissues expressed either undetectable or reduced expression. Aberrant promoter methylation of SOCS-1 was found in 61% of the cervical tumor tissues. SOCS-1 expression and methylation were significantly associated with severity of the disease (p < 0.01). CONCLUSION: We demonstrate for the first time the transcriptional inactivation of SOCS-1 gene due to hypermethylation and synergism with HPV infection which may play an important role in cervical carcinoma.

Microsatellite analysis of genetic population structure of zebu cattle (Bos indicus) breeds from north-western region of India.

The present study aims to understand the existing genetic diversity and structure of six native cattle breeds (Rathi, Tharparkar, Nagori, Mewati, Gir, and Kankrej) adapted to the north-western arid and semi-arid region of India based on microsatellite loci. Various diversity estimates, mean number of alleles (12.84); effective number of alleles (5.02); gene diversity (0.769), and observed heterozygosity (0.667) reflected the existence of substantial within-breed diversity in all the investigated cattle breeds. Mean estimates of F-statistics: F(IT) = 0.144 ± 0.023, F(IS) = 0.071 ± 0.021, and F(ST) = 0.078 ± 0.014 were significantly different from zero (P < 0.05). The interbreed relationships indicated moderate level of breed differentiation between the six cattle breeds with least differentiation between Kankrej-Mewati pair. The phylogeny structuring further supported close grouping of Kankrej and Mewati breeds. Correspondence analysis plotted Rathi, Tharparkar, and Gir individuals into three separate areas of multivariate space; whereas, Kankrej, Mewati, and Nagori cattle showed low breed specific clustering. This reflected the existence of discrete genetic structure for Tharparkar, Rathi, and Gir, the prominent dairy breeds of the region; whereas, admixture was observed for Kankrej, Mewati, and Nagori individuals.

Cocktail of gp63 and Hsp70 induces protection against Leishmania donovani in BALB/c mice.

The 63-kDa antigen of Leishmania donovani is a membrane-anchored matrix metalloprotease that has been shown to be involved in the infection process. We have shown that this antigen alone generates a Th1 type of protective response that is partial but when the animals are primed with the antigen along with the Hsp70, the level of protection is raised significantly, which is demonstrated by a considerable reduction in parasite load of immunized animals when compared to the infected controls. Delayed-type hypersensitivity responses to leishmanin were measured as an index of cell-mediated immune response and were found to be higher in immunized animals when compared to the infected controls, the maximum being in the animals immunized with cocktail of both the antigens. Maximum IgG2a and minimum IgG1 levels were observed in this group of animals. These animals also generated maximum levels of IFN-γ and IL-2 and minimum levels of IL-4 and IL-10 pointing towards the generation of a protective Th1 response and the suppression of the Th2 type of immune response.

Solid lipid nanoparticles regulate functional assortment of mouse mesenchymal stem cells.

A rapid decline in self-renewability, viability and function, of isolated stem cells are major hurdles in developing cell based therapies. There has been an increasing interest towards identifying a support material for maintaining stem cell features of the isolated cells. Pioneering observations of the present paper, demonstrate functionally diverse potential of Solid Lipid Nanoparticles (SLNs) in deciding the fate & behavior of mouse mesenchymal stem cell. The evidences are provided to show the dual nature of the SLNs for being a scaffold for the stem cell attachment, to retain stemness, and as reagent for inducing stem cell differentiation. Scanning electron microscopic examinations together with expression analysis were used to conform to such observations. Results of the study thus suggest that Solid lipid nanoparticles can be used as a good support material when functionalized to achieve adhesive properties and as a molecular paradigm for studying the adipocytic differentiation. We envisage a new role of SLNs towards regulating stem cell character by orchestrating the structural alignment during preparation of Solid lipid nanoparticles.

Insights into the role of IL-12B and IFN-gamma cytokine gene polymorphisms in HIV-1/AIDS infection.

One of the main characteristics of HIV-1 infection is persistent systemic immune activation. This immune activation and dysregulation is characterized by a specific pattern of cytokine production, expression of membrane activation molecules on the cells of the immune system, and changes in the levels of several immune parameters in blood. Therefore, the aim of the present work was to evaluate the effect of a Taq1 polymorphism in the 3'UTR of the IL-12B gene at position -1188 (A/C) and the biallelic polymorphism in the first intron of IFN-gamma at position +874 (T/A) on HIV-1/AIDS among north Indian population. IL-12B and IFN-gamma gene polymorphisms were studied in 300 patients with HIV-1/AIDS and an equal number of negatively diagnosed controls of the matched age, using DNA-based polymerase chain reaction with sequence-specific primers and restriction digestion. The allelic as well as genotypic frequencies of interleukin-12B gene polymorphisms did not significantly differ between HIV-1/AIDS patients and negative healthy controls. A statistically significant correlation was found between IFN-gamma polymorphism and the risk of the disease. The present study suggested that individuals with mutant homozygous IFN-gamma AA genotype were at risk of HIV-1/AIDS (OR = 1.88, 95% CI 1.14-3.10, P = 0.008).

Association of ACE and FACTOR VII gene variability with the risk of coronary heart disease in north Indian population.

The angiotensin converting enzyme (ACE) is a key factor in the production of angiotensin II and in the degradation of bradykinin. Chronic exposure to high levels of circulating and tissue ACE predispose to vascular wall thickening and atherosclerosis. Factor VII (FACTOR VII) is the first enzyme in the extrinsic pathway of the blood coagulation system and plays a key role in hemostasis; it also contributes to the occurrence of thrombotic events. In this study, we have examined the association of ACE and FACTOR VII gene in coronary heart disease patients (n = 300) and their age-matched controls (n = 300). Genotyping was done by PCR-RFLP method. No significant difference was observed in the distribution of I/D genotypes of ACE between cases and controls. In case of FACTOR VII R353Q polymorphism, there was not much difference in the distribution of alleles. AA genotype had protective effect for CHD (OR 0.56, 95% CI 0.37-0.83, P = 0.001). In case of FACTOR VII VNTR, there was difference in the distribution of alleles, H6 (73.5) and H7 (25.5) in cases, and H6 (70.5) and H7 (30.5) in controls. H6H7 and H7H7 genotypes had a protective effect for CHD with OR 0.27, 95% CI 0.18-0.41, P < 0.001, and OR 0.18, 95% CI 0.09-0.36, P < 0.001. Our study showed D allele of ACE to be associated with marginal risk of CHD, AA genotype of FACTOR VII R353Q and H6H7 and H7H7 genotypes of FACTOR VII VNTR showed protective effect for CHD.

VEGF and IL-4 gene variability and its association with the risk of coronary heart disease in north Indian population.

Vascular endothelial growth factor (VEGF) is a potent angiogenic growth factor that has been shown to play a significant role in neovascularization during inflammation in atherosclerotic plaques, formation of collateral vessels to an area of ischemic myocardium and neovascularization at the edges of a myocardial infarction during its repair. Interleukin-4 (IL-4) has important role in immune cell chemotaxis, formation of endothelial cell adhesion molecules and has numerous anti-inflammatory effects which prevent the complications of atherosclerosis, the primary cause of coronary heart disease (CHD). In this study, we have analyzed the effect of 1154 A/G polymorphism of VEGF and 70 bp VNTR polymorphism of intron 3 in IL-4 genes in coronary heart disease (CHD) patients (n = 300) and their age matched controls (n = 300). To analyze polymorphic alleles, ARMS-PCR and RFLP techniques were used. Multiple logistic regression analysis was carried out with statistical software. GG genotype was associated with a decreased risk of development of CHD (OR 0.22, 95% CI 0.12-0.38, P < 0.001). However, A allele showed an increased risk whereas G allele decreased the risk of CHD with diabetes mellitus, hypertension, chronic mental stress and positive familial history of myocardial infarction (MI)/CHD. GG genotype was found to have protective effect with alcohol intake (OR 0.34, 95% CI 0.14-0.82, P < 0.01) and central obesity (OR 0.15, 95% CI 0.04-0.56, P < 0.001). GG genotype of VEGF has also shown significant association with IL-4 (P2P2 and P1P2) genotypes.

The influence of variations in the DNA repair (XRCC1) gene on HIV-1/AIDS among Indian population.

Genetic polymorphisms in DNA repair genes may influence individual variations in the DNA repair capacity. Polymorphisms in the XRCC1 gene that cause amino acid substitutions may impair the interaction of its proteins (XRCC1) with the other enzymatic proteins and consequently alter DNA repair function, which may be associated with the risk of HIV-1/AIDS disease. In this study, we aimed to determine the frequency of polymorphisms in XRCC1 codon 399 in a sample of Indian population with HIV-1/AIDS to evaluate its association with the disease. Polymerase chain reaction and restriction fragment length polymorphism were used to analyse XRCC1 Arg399Gln polymorphisms in 300 positively diagnosed cases with HIV-1/AIDS and an equal number of negatively diagnosed controls of the matched age. The XRCC1 homozygous variant genotype Gln399Gln was associated with an increased risk of HIV-1/AIDS disease (OR = 1.8, 95% CI 1.10-2.94), while no association was found with the Arg399Gln genotype. Polymorphisms in the XRCC1 homozygous variant genotype for the 399Gln allele were associated with the risk of HIV-1/AIDS disease in a sample of North Indian population.

Overexpression of STAT3 in HPV-mediated cervical cancer in a north Indian population.

The constitutively activated STAT family members, particularly STAT3, have been shown to possess transforming properties, and are strongly correlated with tumor development and progression. STAT3 transmits signals from many cytokines and growth factors to target genes in the nucleus through the Jak/Stat signaling pathway. HPV is the main etiological factor in the development of cervical cancer. In the current study, the expression of STAT3 was analyzed in various stages of HPV-mediated cervical carcinogenesis. Tissue biopsies from 100 patients with cervical cancer of different stages and normal tissues from patients undergoing hysterectomy were selected for studying the HPV status and STAT3 expression. HPV status of each corresponding biopsy was analyzed by PCR and typing. The mRNA expression was analyzed by reverse-transcriptase polymerase chain reaction (RT-PCR). HPV infection was detected in majority of cases: 75% (9/12) in precancer, 85% (34/40) stage I & II, and 95% (36/38) in stage III & IV of cervical cancer cases by L1 PCR. Further sub typing revealed HPV16 in 100% (9/9) of L1 positives in precancerous & 90% (63/70) in different stages of cancer. Significant level of STAT3 mRNA expression was predominantly found in cervical cancer cases as compared to normal controls (P = 0.001). We also found a significant correlation of STAT3 expression in cases infected with HPV (P = 0.001). Our results indicate a potentially interactive effect between HPV 16/18 and transcriptional activation of STAT3 gene in cervical carcinogenesis. To our knowledge, this is the first such study to be reported from India. Further investigations are needed to determine the influence of STAT3 expression on cervical carcinogenesis and its possible interaction with HPV infection status.

CpG island methylation of TMS1/ASC and CASP8 genes in cervical cancer.

BACKGROUND: Gene silencing associated with aberrant methylation of promoter region CpG islands is an acquired epigenetic alteration that serves as an alternative to genetic defects in the inactivation of tumor suppressor and other genes in human cancers. AIMS: This study describes the methylation status of TMS1/ASC and CASP8 genes in cervical cancer. We also examined the prevalence of TMS1/ASC and CASP8 genes methylation in cervical cancer tissue and none--neo plastic samples in an effort to correlate with smoking habit and clinicopathological features. METHOD: Target DNA was modified by sodium bisulfite, converting all unmethylated, but not methylated, cytosines to uracil, and subsequently amplified by Methylation Specific (MS) PCR with primers specific for methylated versus unmethylated DNA. The PCR product was detected by gel electrophoresis and combined with the clinical records of patients. RESULTS: The methylation pattern of the TMS1/ASC and CASP8 genes in specimens of cervical cancer and adjacent normal tissues were detected (5/80 (6.2%), 3/80 (3.75%)-2/80 (2.5%), 1/80 (1.2%) respectively). No statistical differences were seen in the extent of differentiation, invasion, pathological type and smoking habit between the methylated and unmethylated tissues (P > 0.05). CONCLUSION: The present study conclude that the frequency of TMS1/ASC and CASP8 genes methylation in cervical cancer are rare (< 6%), and have no any critical role in development of cervical cancer.

Effect of NBS1 gene polymorphism on the risk of cervix carcinoma in a northern Indian population.

Cervical cancer is one of the most common neoplastic diseases affecting women, with a worldwide incidence of almost half a million cases. A history of smoking and use of oral contraceptives have been confirmed to be risk factors for cervical cancer. Genetic susceptibility and immune response, especially impaired cellular immune response, may well be related to the development of cervical cancer. NBS1 is one of the key proteins participating in the recognition and repair of double-strand breaks that may lead to genomic instability and cancer if unrepaired. The objective of the present study was therefore to investigate NBS1 Glu185Gln gene polymorphisms and the risk of cervix cancer in a northern Indian population. We found that passive smokers having particular NBS1 genotypes (Glu/Gln, Gln/Gln or Glu/Gln + Gln/Gln)have an increased risk of developing cervix cancer (OR 5.21, p=0.000001; OR 4.60, p=0.001; OR 5.10, p=0.0000009, respectively).The risk was increased 2.4-fold in oral contraceptive users with a Glu/Gln genotype. We conclude that the risk of cervical cancer is increased in passive smokers and in users of oral contraceptives with certain NBS1 genotypes.

Expression and polimorphism of IFN-gamma gene in patients with cervical cancer.

BACKGROUND: Cervical cancer, the second most common malignancy in women worldwide, is almost invariably associated with infection by human papillomavirus (HPV). However, although many women are infected with high-risk types of HPV, only a subset of infected women will ever develop cervical cancer. Several studies suggested that immunological components play a key role in the development of cervical cancer. Interferon gamma (IFN-gamma ) is a cytokine produced by activated T cells and natural killer (NK) cells that enhances cellular immune responses by increasing T-cell cytotoxicity and NK cell activity. AIM: To study single nucleotide polymorphism (SNP), T to A, located at the +874 position and measure IFN-gamma messenger RNA (mRNA) at the tumor site. METHODS: DNA was isolated from peripheral blood of 200 patients with cervical cancer and 200 healthy controls. The allele polymorphism at position +874 in the IFN-gamma gene was studied by ARMS-PCR (Amplification Refractory Mutation System) and measured IFN-gamma mRNA at the tumor site by means of a semi-quantitative polymerase chain reaction (sqRT-PCR) assay. RESULTS: It was observed that genotypes AT and AA + AT increase the risk of cervical cancer (OR = 3.3, 95% CI - 2.05-5.2, P

Association of Fas-670 gene polymorphism with risk of cervical cancer in North Indian population.

OBJECTIVES: Cervical cancer is the second most common cancer among women in the world, with approximately 470,000 new cases and 231,000 deaths occurring each year. Incidence is greater in developing countries such as India, where this is the most common female malignancy with almost 100,000 new cases each year. Apoptosis must be considered as a safe mechanism that controls the integrity of the cell erasing abnormal clones and it is likely that failure of apoptosis constitutes a key factor responsible for tumor formation, progression and resistance to drugs. The Fas gene plays a key role in regulation of apoptotic cell death and corruption of this signaling pathway has been shown to participate in immune escape and tumorgenesis. STUDY DESIGN: A single-nucleotide polymorphism at -670 of Fas gene promoter (A/G) was examined in a total of 400 blood samples from normal healthy women and cervical cancer patients, using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) technique. RESULTS: Significant association was observed for AG (OR = 3.0, 95% CI = (1.68-5.09, p < 0.001) and combined AG+GG (OR = 2.54, 95% CI = 1.47-4.40, p < 0.001) genotype with risk of cervical cancer. Heterozygous genotype (AG) in SCC showed a highly significant association with risk of cervical cancer (OR = 2.57, 95% CI = 1.47-4.50 p <0.001). Similarly, combined AG+GG genotype had a 2.25-fold risk for SCC patients (OR = 2.25, 95% CI = 1.30-3.90, p < 0.001). There was high increase risk of cervical cancer in passive smokers with AG and combined (AG+GG) genotypes (OR = 4.6, 95% CI = 2.07-10.32, p < 0.001 - OR = 4.9, 95% CI = 2.20-10.32, p < 0.001), respectively. CONCLUSION: This is the first study to provide evidence for the association of a Fas -670 (A/G) gene polymorphism with the risk of cervical cancer in a North Indian population.