RNAi沉默RON基因对人结肠癌HT-29细胞侵袭和耐药性的抑制作用  

Silencing RON gene expression via RNA interference inhibits human colon carcinoma HT-29 cell invasion and drug resistance

作　　者：毛联钢[1] 李克强[1] 卓文莹[2] 戴晓宇[2] 余永明[2] 冯伟云[1] 乐东海[1] 

机构地区：[1]宁波市第二医院肿瘤分子生物学实验室,浙江宁波315010 [2]宁波市第二医院肛肠科,浙江宁波315010

出　　处：《中国肿瘤生物治疗杂志》2012年第5期502-507,共6页Chinese Journal of Cancer Biotherapy

基　　金：宁波市自然科学基金资助项目(No.2008A610082)~~

摘　　要：目的:探讨RNAi沉默酪氨酸激酶受体RON(recepteur d’origine nantais)基因对人结肠癌HT-29细胞侵袭和对抗肿瘤药物敏感性的影响。方法:构建RON基因的RNAi慢病毒载体Lv-RON-siRNA。Real-time PCR和Western blotting检测RON基因的沉默效率及RON蛋白表达水平;Transwell侵袭实验和ATP-TCA(ATP-tumor chemosensitivity assay)检测RON基因对HT-29细胞侵袭和对药物敏感性的影响。结果:慢病毒载体Lv-RON-siRNA感染HT-29细胞对RON基因的沉默效果达到70%。Lv-RON-siRNA感染后,HT-29细胞侵袭力较对照组明显降低(0.97±0.072 vs 1.29±0.076,P<0.05)。Lv-RON-siRNA感染后,HT-29细胞对5-氟尿嘧啶(5-fluorouraci,5-FU)的IC90值和IC50值分别为(14.28±1.34)、(8.93±1.20)μg/ml,顺铂(cisplatin,DDP)的IC90值和IC50值分别为(1.91±0.22)、(0.64±0.07)μg/ml,均明显低于对照组(P<0.01)。结论:沉默RON基因表达能抑制HT-29细胞的侵袭力,提高细胞对5-FU和DDP的敏感性。Objective ： To investigate the effect of receptor tyrosine kinase recepteur d’origine nantais （RON） gene silencing on the invasion and anticancer drug resistance of human colon carcinoma HT-29 cells. Methods： RNAi lentiviral vector targeting RON gene （Lv-RON-siRNA） was constructed. The efficiency of Lv-RON-siRNA on RON gene silence and RON protein level in HT-29 cells were detected by real-time PCR and Western blotting, respectively. The effects of Lv-RON-siRNA on invasion and drug resistance of HT-29 cells were observed by Transwell assay and ATP-TCA （ATP-tumor chemosensitivity assay）. Results： The silencing effect of Lv-RON-siRNA on RON gene expression in HT-29 cells reached 70%. Compared with the control group, the invasion of HT-29 cells in Lv-RON-siRNA infection group was decreased（097±0.07 vs 1.29±0.08, P〈0.05）. The values of IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to 5-FU were （14.28±1.34） μg/ml and （8.93±1.2） μg/ml, respectively. The IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to cisplatin （DDP） were （1.91±0.22） μg/ml and （0.64±0.07） μg/ml, respectively, and were significantly lower than those in the control group （P〈0.01）. Conclusion： Silencing RON gene expression can decrease the invasion ability of HT-29 cells and increase the sensitivity of HT-29 cells to 5-FU and DDP.

关 键 词：RON基因 HT-29细胞 RNAI 侵袭 耐药 

分 类 号：R735.3[医药卫生—肿瘤] R730.54[医药卫生—临床医学]