Study On The Susceptibility Of The Allelic Polymorphism Of ApocⅢ Promoter In Hypertriglyceridemia

Objective: (1)To establish a method of polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) combining agarose gel electrophoresis to analyze the T-455C polymorphism of apoCⅢgene promotor. (2)To establish a method of PCR-RFLP combining microfluidic chip electrophoresis to analyze the C-482T polymorphism of apoCⅢgene promotor. (3)To evaluate the two methods. (4)To analyze the T-455C and C-482T polymorphisms of apolipoprotein CⅢ(apoCⅢ) gene promoter to investigate the association of the two polymorphisms of apoCⅢwith hypertriglyceridemia(HTG) susceptibility.Methods: (1) Polymerase chain reaction(PCR) was performed to amplify apoCⅢincluding T-455C and C-482T polymorphisms, PCR production was digested with BseGI and MspI respectively, and apoCⅢgenotypes were analyzed after agarose gel electrophoresis and microfluidic chip electrophoresis respectively, the levels of serum lipid profiles were also studied by biochemical methods. (2) The polymorphism of apoCⅢgene of research population was analyzed. The apoCⅢgenotypes and alleles frequencies distribution of case control was calculated. (3)To study the relation of apoCⅢgene polymorphism and HTG susceptibility using t-test,χ2 analysis and one-way ANOVA.Results: (1)A method of PCR-RFLP in combination with agarose gel electrophoresis was established to analyze the T-455C polymorphism of apoCⅢgene promotor, which was first time reported in china. (2)Another method of PCR-RFLP in combination with microfluidic chip electrophoresis was established to analyze the C-482T polymorphism of apoCⅢgene promotor, which was first time reported in the world. (3)Three kinds of genotypes detected at apoCⅢ-455 locus were -455TT,-455TC and -455CC. The frequencies of the -455C allele(50.44% vs 33.11%) and of the -455CC genotype(26.55% vs 12.16%) were both significantly different in HTG patients with controls(P < 0.01). Compared with the -455T allele, the -455C allele had a significantly increased HTG risk with odds rations(OR) were 2.06(95%CI: 1.31～3.24). (4)Three kinds of genotypes detected at apoCⅢ-482 locus were -482CC,-482CT and -482TT. The frequencies of the -482T allele(57.14% vs 37.84%) and of the -482TT genotype(38.39% vs 16.22%) of apoCⅢwere both significantly different in HTG patients with controls(P < 0.01). Compared with the -482C allele, the -482T allele had a significantly increased HTG risk with OR was 2.19(95%CI: 1.40～3.43). (5)The frequencies of the genotype and allele of -455 locus and -482 one were both no significantly different in HTG patients and so were in controls.Conclusions: Two methods of PCR-RFLP combining agarose gel electrophoresis and that combining microfluidic chip electrophoresis can investigate the T-455C and C-482T polymorphisms of apoCⅢgene promoter respectively. They are convenient, sensitive, rapid, reliable and suitable for the routine or study work in ordinary laboratory. Both the apoCⅢ-455C allele and -482T one might contribute to an increased risk of HTG among Chinese Han population living in Nantong City.