摘要
目的:探讨下调CACNA2D1对胃癌耐药性细胞系SGC7901/ADR耐药的影响。方法:采用实时荧光定量聚合酶链反应(qRT-PCR)的方法检测16对胃癌及癌旁组织以及胃癌细胞系SGC7901和SGC7901/ADR中的CACNA2D1的mRNA表达水平;蛋白免疫印迹技术(Western blot)检测CACNA2D1的蛋白表达情况;MTT法检测SGC7901、SGC7901/ADR及转染CACNA2D1 siRNA后SGC7901/ADR的IC50值;流式细胞仪检测细胞凋亡。结果:qRT-PCR结果显示:CACNA2D1在胃癌组织中的表达明显高于癌旁组织(P<0.01);CACNA2D1在耐药细胞系SGC7901/ADR中的表达明显高于其亲本细胞系SGC7901(P<0.01)。蛋白免疫印迹技术结果显示相对于转染CACNA2D1 negative control(NC)组,转染CACNA2D1 siRNA组的胃癌细胞CACNA2D1表达降低;MTT结果显示细胞系SGC 7901与SGC7901/ADR的阿霉素半数抑制浓度(IC50)分别为(1.3±0.6)μg/ml与(5.5±0.45)μg/ml;SGC7901/ADR转染CACNA2D1 siRNA后对阿霉素的IC50明显下降。流式细胞术检测凋亡结果显示,CACNA2D1的表达下调后,SGC7901/ADR细胞的凋亡比明显增加。结论:CACNA2D1能够促进胃癌SGC7901/ADR细胞系产生多药耐药。
Objective： To investigate the influence of CACNA2D1 knock - down on drug resistance in gastric cancer cell line SGC7901/ADR. Methods：The mRNA levels of CACNA2D1 in SGC7901 ,SGC7901/ADR and 16 pairs of the gastric cancer tissues and adjacent normal tissues were determined by qRT - PCR assays. The influence of CAC- NA2D1 on the IC50 and apoptosis of cell line SGC7901/ADR were evaluated using MTT and FACS assay. Results： CACNA2D1 mRNA levels were upregulated in gastric cancer tissues compared with adjacent normal tissues （P 〈 0.01 ）, and the mRNA level of CACNA2D1 in SGC7901/ADR was significantly higher than that in SGC 7901 （P 〈 0.01 ）. MTT and FACS assay showed that the muhidrug resistance were reversed by CACNA2D1 knock - down in SGC7901/ADR cells. Conelusion：CACNA2D1 could contribute to the multidrug resistance of gastric cancer cell line SGC7901/ADR.
出处
《现代肿瘤医学》
CAS
2015年第10期1325-1328,共4页
Journal of Modern Oncology
基金
国家自然科学基金资助项目(编号:81301883)
作者简介
王凯（1988一），男，山东潍坊人，医师，硕士研究生，主要从事胃癌的临床与基础研究。E—mail：871883489@qq．com
【通讯作者】樊代明（1953一），男，四川重庆人，中国工程院院士，教授，主任医师，主要从事胃癌的临床与基础研究。E—mail：fandaim@fmmu．edu．cn
