【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。更多还原

【Abstract】 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 IC90 and IC50 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 IC90 and IC50 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.更多还原