Clinical Application Research Of MDR1C3435T Single Nucleotide Polymorphisms In Ovarian Cancer

Objective:To build the method of DNA electrophoresis on horizontal polyaceylamide gels(PAGE) and detect multi-drug resistant gene1 single nucleotide polymorphisms(MDR1SNPs) with the method. To identify the distribution feature of MDR1C3435T gene polymorphism in Hans in ChongQing area by polyacrylamide gel DNA horizontal electrophoresis.To investigate the association of single nucleotide polymorphisms in MDR1C3435T with ovarian cancer.Methods:To investigate the condition of PAGE DNA horizontal electrophoresis with DNA markers.MDR1C3435T gene from normal individuals were amplified with PCR and restricted enzyme digested were done with MboⅠand maps analysis were done with PAGE DNA horizontal electrophoresis. MDR1C3435T gene was also determined by PCR-RFLP method in ovarian cancer patients(cases group) and unrelated healthy women (control group).Results: 10ml of PAGE contains 70μl of 40% Ammonium Persuifate and 20μl of TEMED and prepared the gel in an agarose horizontal electrophoresis tray without air. The efficiency is best with the gel combination rate 29:1 and the concentration of 7.5%. The gene types of MDR1SNPs C3435T in 45 healthful people are: type C/C, 20; T/C, 17; T/T, 8. Phenotype frequency in 251 healthful people: MDR1C3435T T/T0.163, T/C0.470, C/C0.367. Genotype frequency: MDR1C3435T: T0.398 ,C0.602.The frequency of C/C genotype in cases(30.37%)was significantly lower than in controls(47.95%), and the frequency of T/C(44.44%)and T/T(25.19%) in cases was significant respectively higher than in controls (T/C38.35%, T/T13.7%). The frequency of MDR1C3435T null genotype in cases(25.19%)was higher than in controls(13.7%).Compaired with controls,the susceptibility of ovarian cancer with the T/T genotype was increased by 2.121-fold.There showed significant different respectively in frequency of MDR1C3435T null and nonnull genotype in cases and controls in average age(P <0.05). More than 50-year-old age group, the frequency of MDR1C3435T null genotype in cases(20.41%)was higher than in controls(5.1%). Compaired with controls,the susceptibility of ovarian cancer with the T/T genotype was increased by 2.533-fold.Conclusion: 1.It is simple, easy and efficient to detect gene types of MDR1SNPs with PAGE DNA horizontal electrophoresis. It could be recommended using in laboratories lack of resource. 2.The MDR1C3435T gene distribution in Hans in ChongQing close to those in other areas in China ,regional differences not obvious. MDR1C3435T gene mutation genotype could increase risk of ovarian cancer. 3.MDR1C3435T polymorphism is an important risk factor with susceptibility to ovarian cancer.Age increased the risk of ovarian cancer patients will increase.More than 50-year-old age women, the risk of ovarian cancer increased in T/T genotype. 4. The MDR1C3435T gene may be a new target for early screening to ovarian cancer.