Effect of inhibition of hPOT1 expression mediated by siRNA on Caspase-3 in HeLa cells / 中国药理学通报

Aim To inhibit the expression of human protection of telomere gene hPOT1 in HeLa cells using vector based on RNA interference(RNAi) technique and to investigate its effects on the activation of Caspase-3 in HeLa cell.Methods Three recombinant plasmids(our group constructed before) containing different hPOT1 target sequences(pmU6-shRNA1,pmU6-shRNA2 and pmU6-shRNA3)were transfected into HeLa cells by liposomal.The expression inhibition of hPOT1 was detected by RT-PCR and EMSA.The expression and activation of Caspase-3 were inspected by RT-PCR,Western blot analysis and Caspase-3 kit.Results After 48 hour′s transfecting,hPOT1 mRNA and protein level in HeLa cells reduced and the expression of Caspase-3 mRNA had no change.Caspase-3 zymogen decreased but its activity increased.Conclusions hPOT1 expression in HeLa cells can be significantly inhibited by using siRNAvectored RNAi and the down-regulation of hPOT1 expression can activate Caspase-3.

Relationship between single nucleotide polymorphisms of hPOT1 genes and helicobacter pylori infection in gastric carcinoma / 西安交通大学学报(医学版)

Objective To explore the relationship between the single nucleotide polymorphisms(SNPs) of IVS13-98G/T of human protection of telomeres 1(hPOT1) genes and H.pylori infection.Methods A total of 150 cases of gastric carcinoma and 156 cancer-free controls were genotyped by polymerase chain reaction(PCR) and PCR-restriction fragment length polymorphism(PCR-RFLP) method.H.pylori was identified with PCR and Warthin-Starry methods.Results The frequencies of GG,GT,and TT genotypes of hPOT1IVS13-98 G/T were 22.00%,41.67% and 36.67% in patients and 24.36%,51.92%,and 23.72% in controls,respectively.The OR for any GT and GG genotype was 0.432(95% CI: 0.242-0.772,P=0.005) and 0.540(95% CI: 0.274-1.064,P=0.075) when compared with TT genotype,respectively.There were no differences in genotype distribution and allele frequency of IVS13-98 G/T polymorphism(?2=3.0257,P= 0.0820).hPOT1IVS13-98 G/T polymorphism was not correlated with H.pylori infection in gastric cancer.Conclusion Our results indicated that IVS13-98 G/T SNP of hPOT1 gene might be associated with reduced risk for gastric carcinoma,but not with H.pylori infection.