Linkage analysis between prostate cancer occurrence and Y-chromosomal DYS loci in Malaysian male subjects

Prostate cancer has become the second leading cancer among men across ethnic groups in the world. Since it is influenced by a complex genetics that may affect the level of susceptibility for the development of the disease, four Y-linked short tandem repeats (STRs), DYS388, DYS435, DYS437, and DYS439 were genotyped to compare Malaysian prostate cancer patients and normal controls males. A total of 175 subjects comprising 84 patients and 91 healthy individuals from three major ethnics were recruited. Multiplex PCR was optimized to co-amplify all four DYS loci. All samples were genotyped for alleles of four DYS loci using a Genetic Analysis System. Result showed that allele 10 (A) of DYS388 had a significantly lower incidence towards disease than other alleles of this locus, while allele 12 (C) of DYS388 and allele 14 (E) of DYS439 showed a significantly higher risk to develop prostate cancer compared to other alleles of these loci. Moreover, among 47 different haplotypes comprising different alleles of four DYS loci found in the overall study samples, it is noticed that AABC and CAAA showed a lower and higher frequency among cases than controls, respectively. As a conclusion, Malaysian males who belong to Y-lineages with either allele 12 of DYS388, allele 14 of DYS439, or haplotype CAAA tend to develop prostate cancer. Meanwhile, those belonging to Y-lineages with allele 10 of DYS388 or haplotype AABC are more resistant to the disease. Thus, it is suggested that genetic elements give an influence on the development of prostate cancer

Linkage between prostate cancer occurrence and Y-chromosomal DYS loci in Malaysian subjects..

Purpose: Prostate cancer differs markedly in incidence across ethnic groups. Since this disease is influenced by complex genetics, it is many genetic factors may affect the level of susceptibility to development of the disease. In this study, four Y-linked short tandem repeats (STRs), DYS388, DYS435, DYS437, and DYS439, were genotyped to compare Malaysian prostate cancer patients and normal control males. Materials and methods: A total of 175 subjects comprising 84 patients and 91 healthy individuals were recruited. Multiplex PCR was optimized to co-amplify DYS388, DYS435, DYS437, and DYS439 loci. All samples were genotyped for alleles of four DYS loci using a Genetic Analysis System. Results: Of all DYS loci, allele 10 (A) of DYS388 had a significantly lower incidence of disease in compare with other alleles of this locus, while a higher incidence of disease was found among males who had either allele 12 (C) of DYS388 or allele 14 (E) of DYS439. Moreover, a total of 47 different haplotypes comprising different alleles of four DYS loci were found among the whole study samples, of which haplotypes AABC and CAAA showed a lower and higher frequency among cases than controls, respectively. Conclusions: It is likely that Malaysian males who belong to Y-lineages with either allele 12 of DYS388, allele 14 of DYS439, or haplotype CAAA are more susceptible to develop prostate cancer, while those belonging to lineages with allele 10 of DYS388 or haplotype AABC are more resistant to the disease

Determining Correlation Between Males from Different y-Chromosomal Lineages and their Susceptibility Resistance to Prostate Cancer Using Multiple Dys and Binary Markers

Prostate cancer is a major health burden throughout the world. It is proposed that a complex genetics influences on prostate cancer and a similarity in genetic elements may affect the susceptibility or resistance of men to this disease. Since the geographical specificity of haplogroups implies that males from different ethnic groups have various Y-lineages with different Y-chromosomal haracteristics, using of Y-chromosomal markers (such as DYS and Binary markers) might be a powerful tool to investigate the susceptibility or resistance of men to prostate cancer. In this study, four Y-linked polymorphic microsatellite markers including DYS388, DYS435, DYS437, and DYS439, and 10 Y-linked binary markers including M20, M35, M96, M120, M172, M201, M207, M217, M231, and M258 were examined to classify Malaysian and Iranian prostate cancer patients and normal control males residing in both countries. The study group included a total of 373 subjects comprising 84 samples from Malaysian prostate cancer patients (36 Malays, 44 Chinese, and 4 Indians), 91 samples from Malaysian healthy control group (40 Malaysian-Malays, 47 Malaysian- Chinese, and 4 Malaysian-Indians), 98 samples from Iranian prostate cancer patients,and 100 samples from Iranian healthy control group. The peripheral blood was collected from all participants for genomic DNA extraction. For DYS markers, a multiplex PCR was optimized to co-amplify four above mentioned DYS loci in a single reaction tube. All DNA samples were genotyped for alleles of four DYS loci using a Genetic Analysis System CEQ8000 (Beckman Coulter). Also, an Allele Specific PCR method was optimized for every above mentioned Y-linked binary loci. In this study,alleles 12 (D) of DYS388 [(Iranians: OR, 3.242; 95% CI, 1.808 – 5.815; P < 0.01) (Malaysians: OR, 3.587; 95% CI, 1.847 – 6.966; P < 0.01) (Malaysian-Malays: OR,2.714; 95% CI, 1.022 – 7.209; P = 0.042) (Malaysian Chinese: OR, 6.006; 95% CI,2.231 – 16.171; P < 0.01) (whole study population: OR, 3.314; 95% CI, 2.147 – 5.116;P < 0.01)] and allele 14 of DYS439 [(Malaysians: P = 0.018 for Malaysians)(Malaysian-Malays: P = 0.030) (whole study population: OR, 8.736; 95% CI, 1.082 – 70.552; P = 0.015)] were significantly associated with a higher risk to develop prostate cancer. On the other hand, allele 10 (B) [(Malaysians: OR, 0.129; 95% CI, 0.047 – 0.357; P < 0.01) (Malaysian Malays: OR, 0.176; 95% CI, 0.036 – 0.870; P = 0.02)(Malaysian Chinese: OR, 0.070; 95% CI, 0.015 – 0.325; P < 0.01) (whole study population: OR, 0.152; 95% CI, 0.057 – 0.400; P < 0.01)] and 13 (E) of DYS388 [(Iranians: OR, 0.219; 95% CI, 0.09 – 0.533; P < 0.01) (whole study population: OR,0.430; 95% CI, 0.221 – 0.838; P = 0.011) were significantly associated with the lower risk of developing prostate cancer. These findings support the hypothesis that males from different Y-chromosomal origins are different regarding their susceptibility or resistance to prostate cancer. Also, A total of 76 different haplotypes comprising different alleles of four DYS loci were found among the whole study samples, of which haplotypes DABB (Iranians: OR, 7.615; 95% CI, 0.919 – 63.099; P = 0.028), DABD (Iranians: P = 0.022), DAAB [(Malaysian: P = 0.035) (whole study population: OR, 2.862; 95% CI, 0.999 – 8.195; P = 0.042)], DABC (Malaysian-Chinese: P = 0.035),and DAAC (whole study population: OR, 2.134; 95% CI, 0.998 – 4.563; P = 0.046)showed a higher frequency among patients than healthy controls, while haplotypes BABD [(Malaysian: OR, 7.615; 95% CI, 0.042 – 0.930; P = 0.024) (Malaysian-Chinese: OR, 0.201; 95% CI, 0.043 – 0.931; P = 0.024) (whole study populations: OR, 0.201; 95% CI, 0.043 – 0.931; P = 0.024)] and DAAF (whole study population: P =0.021) had a lower frequency among patient than controls. Therefore, it is likely that men who belong to the lineages with DAAB, DABC, DABD, or DABB haplotypes have a higher risk to develop prostate cancer, while those belonging to lineages with either BABD or DAAF haplotypes are less exposed to this disease than other males.However, the evidences do not strongly support the hypothesis regarding the association of DYS haplotypes with the risk of prostate cancer. The comparison between frequencies of 10 haplogroups studied in this research showed no significant differences between prostate cancer patient and healthy control groups in all populations studied. In conclusion, results of this study emphasize the influence of genetic elements on prostate cancer and some alleles of DYS388 and less likely DYS439 as well as some DYS haplotypes have the potential to be used as a screening method for prediction of susceptibility to prostate cancer both in Malaysian and Iranian populations

Using High Resolution Melting method for detecting nucleotide polymorphisms of STK11 gene germline in patients with digestive system cancers

Abstract Background and Objective: High resolution melting (HRM) analysis is a technique to measure decrease of fluoresce, caused by discharge of the dye, throughout DNA thermal melting gradient process. STK11 gene encodes one of cellular serine-threonine kinase proteins that regulates cellular polarity. It also acts as tumor suppressor protein. Mutations of germline in the gene is coincident with Peutz-Jeghers syndrome and potential of developing a variety of neoplasia. Methodology: In an experimental study, genome DNA of 56 patients with digestive system cancer was extracted. Afterward, nucleotide changes over the gene were examined using Real-time PCR and High resolution melting (HRM). Findings: Nucleotide screening using HRM technique revealed two types of SNP in introns 6 & 7 in 10 patients. Four patients showed homozygous C/T nucleotide changes [cluster id/dsSNP/rs9282860] in intron 6 and six patients showed heterozygous C/G nucleotide changes [cluster id/dsSNP/rs2075607] in intron 7. Comparison of HRM results with sequencing results indicated 100% conformity. Conclusion: Although no mutation was observed in exon section of the gene, primary screening of STK11 gene to diagnose unknown nucleotide changes of germline and somatic in patients with neoplasia using HRM was feasible, easy, and cost effective