Genotype Combination Distribution And Frequency Distribution Of SNP At Three Sites Of KLOTHO Gene In Elderly People Of Chinese Han Nationality Population In Chongqing And The Association Of The Polymorphisms With Primary Osteoporosis

Objective1. To study genotype combination distribution and allele frequency distribution of single nucleotide polymorphisms (SNP) at three sites of Klotho gene of G-395A,F352V and C370S in elderly people of Han nationality in Chongqing.2. To study the association between primary osteoporosis and single nucleotide polymorphisms (SNP) at three sites of Klotho gene of G-395A, F352V and C370S in elderly people of Han nationality in Chongqing.Methods1. The SNPs of Klotho G-395A, F352V and C370S were detected by allelic specific primer(ASP)polymerase chain reaction(PCR).The genotypes combination distribution of SNPs at three sites of Klotho was determined by clustering analysis technique. 2. The G-395A,F352V and C370S single nucleotide polymorphism of Klotho gene were tested by allelic specific primer(ASP) polymerase chain reaction(PCR)in 300 patients with primary osteoporosis and 93 healthy elderly people of Han nationality in Chongqing. The distribution frequency of genotypes was compared each other in the two groups as well as in the gender subgroups.Results1. The genotype frequencies of GG,GA and AA were 47.69%,42.61% and 9.70% respectively in the Klotho G-395A SNP. G and A allele haplotype frequencies of Klotho G-395A SNP was 69.00% and 31.00% respectively. The genotype frequencies of FF,FV and VV were 35.52%,58.93% and 5.55% respectively in the Klotho F352V SNP. F and V allele haplotype frequencies of Klotho F352V SNP was 64.98% and 35.02% respectively. The genotype frequencies of CC,CS and SS were 37.32%,58.80% and 3.88% respectively in the Klotho C370S SNP. C and S allele haplotype frequencies was 66.72% and 33.28% respectively in the Klotho C370S SNP. The preceding 10 combined genotypes in the natural combination distribution of three sites of SNPs of Klotho gene were ranked as follows: 253 subjects with combined genotype of Klotho G-395G+F352V+C370S(16.92%),241 subjects with combined genotype of Klotho G-395A+F352V+C370S(16.12%),152 subjects with combined genotype of Klotho G-395G+F352V+C370C(10.17%),131 subjects with combined genotype of Klotho G-395A+F352V+C370C(8.76%),126 subjects with combined genotype of Klotho G-395G+F352F+ C370S(8.43%),121 subjects with combined genotype of Klotho G-395A+F352F+C370S(8.09%),109 subjects with combined genotype of Klotho G-395G+F352F+C370C(7.29%),86 subjects with combined genotype of Klotho G-395A+F352F+C70C(5.75%), 52 subjects with combined genotype of Klotho A-395A+F352V+C370S(3.48%),30 subjects with combined genotype of Klotho A-395A+F352F+C370C( 2.01%),30 subjects with combined genotype of Klotho G-395G+V352V+ C370S(2.01%).2. The frequency of AA genotypes of Klotho G-395A in the osteoporosis group was significantly higher than that in the control group (P<0.01). According to gender stratification analysis, only the distribution frequency of AA genotypes of Klotho G-395A in male osteoporosis subgroup was significantly higher than that in the male control subgroup(P<0.01). There was no significant difference to be found in the frequency of FF,FV or VV genotypes of Klotho F352V either in the male or female subgroup between the osteoporosis group and the control group(P>0.05). There was no significant difference to be found in the frequency of CC,CS or SS genotypes of Klotho C370S either in male or female subgroup between the osteoporosis group and the control group (P>0.05). Conclusions1. This study shows the features of combination distribution and frequency distribution of Klotho SNPs and provides clues and evidences for further study of Klotho gene and its expression.2. The G-395A SNP of Klotho gene is associated with primary osteoporosis,which has sex difference between the two groups. No association of SNPs of the F352V and C370S of Klotho gene with primary osteoporosis has been found.