Kamlesh Guleria

p.R72P, PIN3 Ins16bp Polymorphisms of TP53 and CCR5Δ32 in North Indian Breast Cancer Patients

BACKGROUND The present study aimed to find the prognostic implications of two polymorphisms in TP53 (p.R72P, PIN3 Ins16bp) and one in CCR5 (CCR5/32) in sporadic breast cancer patients. METHODS DNA samples of 80 breast cancer patients and 80 age and gender matched unrelated healthy control individuals from Punjab, North West India were analyzed. RESULTS For p.R72P, the genotype frequency was 13.8% (RR), 58.8% (RP), 27.5% (PP) in patients and 33.9% (RR), 40.0% (RP), 26.5% (PP) in controls. For PIN3 Ins16bp, the genotype frequencies were 53.75% (A1A1), 37.5% (A1A2), 8.75% (A2A2) in patients and 66.3% (A1A1), 31.3% (A1A2), 2.5% (A2A2) in controls. Only 4 (5%) breast cancer patients were heterozygous for CCR5Δ32 deletion. Common RR-A1A1-WT/WT genotype was lower while RP-A1A2-WT/WT genotype was higher in patients as compared to controls. RP-A1A1-WT/WT genotype was significantly higher in patients as compared to control individuals (p = 0.008). CONCLUSION Though a clear association of any particular genotype with sporadic breast cancer or stage was not apparent, the results of present study were suggestive that sporadic breast cancer patients with RR-A1A1-WT/WT genotype might have a better response to chemotherapy, thus improving their chances of survival.

ABO Blood Groups in Gastrointestinal Tract (GIT) and Breast Carcinoma Patients

Abstract The ABO blood group distribution varies in different geographical and ethnic groups. In Punjab (India), the ABOblood frequency is B>O>A>AB. The current study was an attempt to correlate the ABO blood group frequency with preponderanceof breast and gastrointestinal cancer in Punjab, to assess the utility of ABO blood group as a preclinical marker. The study sampleconsists of 160 cancer patients and 160 controls. In cancer patients, the incidence of A and B groups was significantly highercompared to controls. In breast cancer, the frequency of A group was significantly higher and in oesophageal cancer, the frequencyof B group was significantly higher. The results indicate that blood type should be considered along with other risk factors tounderstand the individual patient’s risk.

TP53 polymorphisms in sporadic North Indian breast cancer patients.

BACKGROUND The purpose of this study was to evaluate the potential association of five (p.P47S, p.R72P, PIN3 Ins16bp, p.R213R and r.13494g>a) polymorphisms of TP53 with the risk of developing breast cancer in North Indian Punjabi population. METHODS We screened DNA samples of 200 sporadic breast cancer patients (197 females and 3 males) and 200 unrelated healthy, gender and age matched individuals for the polymorphisms. RESULTS For the p.P47S polymorphism, we observed the PP genotype in 99.5% of the patients and PS genotype in only 1 patient. All the controls had the wild type PP genotype. The frequency of RR, RP and PP genotype of p.R72P was 23.5% vs 33.5%, 51.5% vs 45.5% and 25% vs 21% in patients and controls respectively. Heterozygous (RP) genotype was increased in breast cancer patients as compared to controls (51.5 vs 45.5%) and showed 1.61 fold significantly increased risk for breast cancer (OR=1.61, 95% CI, 1.01-2.58, p=0.04). In breast cancer patients the frequencies of A1A1, A1A2 and A2A2 genotypes of PIN3 Ins16bp polymorphism were 67%, 26% and 7% respectively whereas in controls the genotype frequencies were 68.5%, 27.5% and 4% respectively, with no significant difference. For p.R213R (c.639A>G), all individuals had homozygous wild type genotype. The frequencies of GG, GA and AA genotypes of TP53 r.13494g>a polymorphism were 62 vs 67.5%, 33 vs 28% and 5 vs 4.5% in patients and controls respectively, again without significant difference. We observed that RP- A1A1 genotype combination of p.R72P and PIN3 Ins16bp and RP-GG combination of p.R72P and r.13494g>a polymorphism showed significant risk of breast cancer (OR=1.65, 95%CI: 0.98-2.78, p=0.05; OR=1.72, 95%CI: 1.01-2.92, p=0.04). CONCLUSION The results of present study indicated that among the five TP53 polymorphisms investigated, the p.R72P polymorphism, and the RP-A1A1 and RP-GG genotype combination contribute to breast cancer susceptibility in North Indians.

Non-random Chromosomal Aberrations in Peripheral Blood Leucocytes of Gastrointestinal Tract and Breast Cancer Patients

Abstract Chromosomal instability was studied in cultured peripheral blood leucocytes to assess whether peripheral blood had non-random cytogenetic aberrations as observed in tumor tissue. The study was conducted on sporadic breast and gastrointestinal tract cancer patients from an area having increased incidence of these cancers. The study sample was of 38 sporadic (26 gastrointestinal tract and 12 breast) cancer patients and 30 controls subjects. Cancer patients had significantly increased (20-68%) aberrant metaphases compared to controls (6.2%). In patients, the aberrations seen were loss or gain of chromosomes, polyploidy, chromatid breaks and gaps, acentric fragments, marker chromosomes, double minutes and acrocentric associations. In oesophageal cancer loss of chromosome 2, 7q-, 10, 11, 12, 15, 17, 19, 21,and Y and gain of chromosome 3, 4, 10, 19, and 22; in gastric cancer loss of 11q and X, gain of extra C group like marker chromosome and in breast cancer loss of 1,2, 5, 7, 11, 12, 13, 15, 16, 18, 22, and X and gain of chromosome 2q, 13, 19 and 20 were seen. Chromatid breaks were seen on chromosomes 1p, 2p, 2q and 4q while chromatid gaps were on chromosomes 1p, 2p, 3p, and 3q only. Aberrations involving specific chromosomes i.e. 2, 7, 11, 12, 15, 19, 22 and X in lymphocytes of cancer patients having cancers of diverse sites indicate that the patients probably have a constitutional chromosomal instability which participates in cancer predisposition and there is involvement of some common genes in tumor initiation and development.

No Association of Hypoxia Inducible Factor-1α Gene Polymorphisms with Breast Cancer in North-West Indians

BACKGROUND Hypoxia inducible factor-1 alpha (HIF-1α) is the key regulator of cellular responses to hypoxia and plays a central role in tumour growth. Presence of Single nucleotide polymorphisms (SNPs) in the critical regulatory domains of HIF-1α may result in the overexpression of the protein and subsequent changes in the expression of the downstream target genes. The aim of study was to investigate the association of three SNPs (g.C111A, g.C1772T and g.G1790A) of HIF-1α with the risk of breast cancer in North Indian sporadic breast cancer patients. MATERIALS AND METHODS A total of 400 subjects, including 200 healthy controls and 200 patients with breast cancer were recruited in this study. Genotypes were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS The CC and CA genotype frequency of HIF-1α g.C111A polymorphism was 100 vs 99% and 0 vs 1% in breast cancer patients and healthy controls respectively. The frequencies of CC, CT and TT genotype of g.C1772T polymorphism were 76 vs 74.5%, 19 vs 21% and 5 vs 4.5% in breast cancer patients and control individuals respectively. There was no significant difference in genotype and allele frequencies of HIF-1α g.C1772T polymorphism between cases and control individuals (p>0.05). For g.G1790A genotypes, all patients and controls had only GG genotype. CONCLUSIONS The three HIF-1α polymorphisms (g.C111A, g.C1772T and g.G1790A) are not associated with breast cancer risk in North-West Indian patients.

Association of −2549 Insertion/Deletion Polymorphism of Vascular Endothelial Growth Factor with Breast Cancer in North Indian Patients

AIMS The aim of the present study was to assess the role of the vascular endothelial growth factor (VEGF) -2549 insertion/deletion (I/D) polymorphism in susceptibility to sporadic breast cancer. METHODS DNA samples of 94 breast cancer patients and 94 unrelated healthy control individuals with matched age and gender from the same geographical region of Punjab, North West India were screened for the -2549 I/D polymorphism. Serum VEGF-C (sVEGF-C) levels of breast cancer patients and healthy controls were measured using an enzyme-linked immunosorbent assay. RESULTS The frequency of the II, ID, and DD genotype was 23.40 versus 10.64%, 48.94 versus 52.13%, and 27.66 versus 37.23%, in patients and controls, respectively. A statistically significant difference was observed for genotype distribution among the patients and controls (χ(2)=6.039, p=0.049). There was a significant increase in the I allele frequency in the patients as compared with controls (47.86 versus 36.70%, p=0.028). The sVEGF-C levels were also considerably higher in patients as compared to healthy controls (p<0.01). CONCLUSIONS The VEGF -2549 I/D polymorphism has a role in the susceptibility to breast cancer in the Amritsar region of Punjab, India.

Association of +405C>G and +936C>T polymorphisms of the vascular endothelial growth factor gene with sporadic breast cancer in North Indians.

BACKGROUND Vascular endothelial growth factor (VEGF), an endothelial cell specific mitogen, has been implicated as a critical factor influencing tumor related angiogenesis. The aim of present study was to evaluate the relationship between VEGF +936C>T and +405C>G polymorphisms of VEGF with risk of breast cancer in Punjab, India. MATERIALS AND METHODS We screened DNA samples of 192 sporadic breast cancer patients and 192 unrelated healthy, gender and age matched control individuals for VEGF +936C>T and +405C>G polymorphisms using the PCR-RFLP method. RESULTS For the VEGF +405C>G polymorphism, we observed significantly increased frequency of GG genotype in cases as compared to controls and strong association of +405GG genotype was observed with three fold risk for breast cancer (OR=3.07; 95%CI 1.41-6.65; p=0.003). For the +936C>T polymorphism, significant associations of CT and combined CT+TT genotypes were observed with elevated risk of breast cancer (p=0.021; 0.023). The combined genotype combinations of GG-CC and GG- CT of +405C>G and +936C>T polymorphisms were found to be significantly associated with increased risk of breast cancer (p=0.04; 0.0064). CONCLUSIONS The findings of the present study indicated significant associations of VEGF +936C>T and +405C>G polymorphisms with increased breast cancer risk in patients from Punjab, North India.

Association of the -2518 A/G Polymorphism of MCP-1 with Breast Cancer in Punjab, North-West India.

BACKGROUND Monocyte chemoattractant protein-1 (MCP-1) is a major chemokine thought to be responsible for monocyte and T-lymphocyte recruitment in acute inflammatory conditions and recruitment of macrophages in tumors. It is also implicated in cardiovascular disease, rheumatoid arthritis and chronic obstructive pulmonary disease. The aim of the present study was to investigate the correlation between MCP-1 -2518 A/G polymorphism and breast cancer risk in patients from Amritsar city of Punjab state in North-West India. MATERIALS AND METHODS We screened DNA samples of 200 sporadic breast cancer patients and 200 age and gender matched unrelated healthy individuals for MCP-1 -2518 A/G polymorphism using the PCR-RFLP method. RESULTS A significantly increased frequency of the GG genotype was observed in patients as compared to controls. Individuals carrying the MCP1 -2518GG genotype had a two fold risk for breast cancer (OR=2.06, 95%CI, 1.06-3.98; p=0.03). Genetic models analysis revealed a significant association between MCP-1 -2518 A/G polymorphism and cancer risk in homozygous co-dominant (OR=2.06, 95%CI, 1.06-3.98; p=0.03) and recessive (OR=1.97, 95%CI, 1.05-3.70; p=0.03) models. CONCLUSIONS We conclude that the GG genotype of the MCP-1-2518 A/G polymorphism is associated with increased risk to breast cancer in Punjab, North-West India.

Analysis of TP53 polymorphisms in North Indian sporadic esophageal cancer patients.

BACKGROUND To investigate the relationship of five TP53 polymorphisms (p.P47S, p.R72P, PIN3 ins16bp, p.R213R and r.13494g>a) with the esophageal cancer (EC) risk in North Indians. MATERIALS AND METHODS Genotyping of p.P47S, p.R72P, PIN3 ins16bp, p.R213R and r.13494g>a polymorphisms of TP53 in 136 sporadic EC patients and 136 controls using polymerase chain reaction and PCR-RFLP. RESULTS The frequencies of genotype RR, RP and PP of p.R72P polymorphism were 16.91 vs 26.47%, 58.82 vs 49.27% and 24.27 vs 24.27% among patients and controls respectively. We observed significantly increased frequency of RP genotype in cases as compared to controls (OR=1.87, 95% CI, 1.01-3.46, p=0.05). The frequencies of genotype A1A1, A1A2 and A2A2 of PIN3 ins16bp polymorphism were 69.12 vs 70.59%, 27.20 vs 25% and 3.68 vs 4.41% among patients and controls. There was no significant difference among genotype and allele distribution between patients and controls. The frequencies of genotype GG, GA and AA of r.13494g>a polymorphism were 62.50 vs 64.70%, 34.56 vs 30.15% and 2.94 vs 5.15% among patients and controls respectively. No significant difference between genotype and allele frequency was observed in the patients and controls. For p.P47S and p.R213R polymorphisms, all the cases and controls had homozygous wild type genotype. The RP-A1A1-GG genotype combination shows significant risk for EC (OR=2.01, 95%CI: 1.01-3.99, p=0.05). CONCLUSIONS Among the five TP53 polymorphisms investigated, only p.R72P polymorphism may contributes to EC susceptibility.

Spectrum of Chromosomal Aberrations in Peripheral Blood Lymphocytes of Gastrointestinal Tract (GIT) and Breast Cancer Patients

Abstract The aim of present study was to assess the spectrum of chromosomal aberrations in peripheral blood lymphocytes of sporadic Gastrointestinal tract (GIT) and Breast cancer patients. Ninety eight patients (56 GIT cancer and 42 breast cancer) and seventy seven unrelated healthy set of control individuals were investigated in the present study. Lymphocytes were cultured using standard protocol. In each case, 100 metaphases were screened for numerical as well as structural aberrations. Higher frequency of aberrant metaphases with chromosomal aberrations including gaps, breaks, terminal deletions, acentric fragments, double minutes, acrocentric associations, premature chromatid separations, pulverisations, polyploidy, loss and gain of chromosomes, ring chromosome and marker chromosomes were observed in cancer patients as compared to controls. A non-random involvement in aberrations of chromosomes harbouring genes implicated in tumorigenesis was observed in GIT as well as in breast cancer patients. Aberrations in peripheral blood lymphocytes (PBLs) can indicate the constitutional anomalies and understanding of molecular basis of chromosomal instability (CIN) phenotype can help in earlier diagnosis or prognosis.