抑制miR-185对人胃癌细胞株SGC7901药物敏感性影响的机制探讨  

Effect and mechanism of miR-185 inhibition to drug resistance of human gastric cancer cell line SGC7901

作　　者：李勇[1] 檀碧波[1] 赵群[1] 范立侨[1] 王冬[1] 刘羽[1] 刘庆伟[1] 

机构地区：[1]河北医科大学第四医院外三科,河北石家庄050011

出　　处：《中华肿瘤防治杂志》2015年第17期1347-1351,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81072033;81372580);河北省自然科学基金(C2010000619);河北省普通高校强势特色学科资助项目(冀教高[2005]52);河北省科技支撑项目(14277779D);河北省卫生厅重大医学科研课题(zd2013040)

摘　　要：目的探讨微小RNA185(microRNA185,miR-185)对人胃癌细胞株SGC7901药物敏感性的影响及相关机制。方法选取2014-01-04-2014-05-20于河北医科大学第四医院住院手术,且经病理确诊的胃腺癌组织标本25例。蛋白质印迹技术检测胃癌组织、癌旁组织及细胞株SGC7901、耐药细胞株SGC7901/ADR和胃上皮细胞株GES-1中miR-185的表达。将miR-185抑制物(Anti-miR-185)转染SGC7901;MTT法检测转染前后肿瘤细胞对化疗药物多柔比星(adriamycin,ADR)、氟尿嘧啶(5-fluorouracil,5-FU)和奥沙利铂(oxaliplatin,L-OHP)的敏感性;荧光定量PCR和蛋白质印迹技术检测转染前后胃癌细胞多药耐药基因1(multidrug resistance gene 1,MDR1)、多药耐药相关蛋白-1(multidrug resistance associated protein-1,MRP-1)、谷胱甘肽S-转移酶-π(glutathione S-transferase-π,GST-π)、拓扑异构酶-Ⅱ(topoisomerase-Ⅱ,TOPO-Ⅱ)和胸苷合成酶(thymidylate synthase,TS)的表达变化。结果miR-185在胃癌组织表达(0.243±0.045)低于癌旁组织(0.910±0.142),t=-22.389,P<0.001;miR-185在SGC7901中相对表达强度为0.630±0.101,在SGC7901/ADR为0.191±0.030,在GES-1中表达为0.903±0.117,三者比较miR-185在SGC7901/ADR中表达最低,P<0.001。Anti-miR-185转染后,SGC7901对ADR、5-FU和L-OHP的敏感性明显减弱,P值均<0.05。Anti-miR-185转染后,SGC7901中MDR1、MRP-1和GST-π水平均较转染前增高,P值均<0.05;而TOPO-Ⅱ和TS水平则无明显变化,P>0.05。结论抑制胃癌细胞中miR-185表达可上调多药耐药基因MDR1、MRP-1和GST-π表达,导致胃癌细胞耐药性增强。OBJECTIVE To explore the effect and mechanism of miR-185 inhibition to drug resistance of gastric cancer cell line SGC7901. METHODS Specimens of 25 patients with gastric carcinoma were enrolled in Fourth Affiliated Hospital, Hebei Medical University from 2014-01-04 to 2014-05-20. The expressions of miR-185 in gastric cancer tis- sues, tumor adjacent tissues and SGC7901, drug-resistant cell line SGC7901/ADR, gastric epithelial cell line GES-1 were tested with Western blot. Anti-miR-185 was synthesized and tansfected into SGC7901. Sensitivity to chemotherapeutic drugs（ADR, 5-FU, L-OHP） of SGC7901 were measured with MTT assay. The expressions of multidrug resistance （MDR） related genes MDR1/P-gp, MRP-1, GST-π, TOPO- Ⅱ , TS were determined by fluorescent quantitation RT-PCR and Western blot. RESULTS Expressions of miR-185 were lower in gastric cancer tissues（0. 243±0. 045） than that in tumor adjacent tissues（0. 910±0. 142;t=-22. 389, P〈0. 001）; and relative expressions of miR-185 in SGC7901 was （0. 630±0. 101）, in SGC7901/ADR was （0. 191±0. 030） and in GES-1 was （0. 903±0. 117）, the lowest expressions of miR-185 was found in SGC7901/ADR（P（0. 001）, Sensitivity to chemotherapeutic drugs of SGC7901 was obviously weakened after Anti-miR-185 tansfected with MTT assay（P〈0.05）. The expression of MDR1/P-gp, MRP-1, GST-π decreased after Anti-miR-185 tansfected （P〈0.05）, and no obvious change was found in TOPO-Ⅱ , TS（P〉0. 05）.CONCLUSION Inhibiting miR-185 can improve resistance of gastric cancer cells to chemotherapeutic drugs by upregulating MDR genes MDR1/P-gp, MRP-1, GST-π.

关 键 词：胃癌 微小RNA-185 多药耐药性 体外药敏实验 

分 类 号：R735.2[医药卫生—肿瘤]