Establishment And Application Of High Resolution Melting Method For Detecting SNP In MDR1 Gene

Objective: It is difficult to cure gliomas completely with surgery. As a widely used methylating agent, temozolomide(TMZ)possesses effective clinical activity against gliomas. However, clinical trials showed that the effectiveness of TMZ was only 45% when used in clinic therapy because of TMZ resistance, which has become a major problem of gliomas chemotherapy. A SNP (C1236T) in MDR1 exon 12 has been found as a new independent prediction factor. Because the previous genotyped methods were time-consuming, tedious, and high cost, we designed to set up a new specificity, sensitivity, but simple, rapid and low-cost genotyped method and to investigate feasibility for detecting TMZ resistance of gliomas chemotherapy.Methods: Firstly,genotyping method to C1236T by PCR-HRM was established with 9 samples of healthy volunteers'genomic DNA, and the results was compared with sequencing. Then,163 samples were genotyped by established PCR-HRM to get C1236T frequency in Chinese population.At last, 26 cases of glioma patients were genotyped directly.Results: The genotyping results with PCR-HRM and sequencing were consistent,and C/C,C/T,T/T frequency of C1236T in Chinese population are 9.2%,44.8% and 46.0% ,C/C,C/T,T/T cases of 26 glioma patients were 6,12 and 8 respectively.Conclusion: PCR-HRM was a high throughput, low-cost, rapid and simple genotype method, which might play a role in checking TMZ resistance of gliomas chemotherapy.