RNA干扰联合阻断MMS2和REV3基因表达减轻结肠癌细胞系THC8307/L-OHP对奥沙利铂的耐药性  

Interference of MMS2 and REV3 gene by RNA reduces the Oxaliplatin resistance of human colon-carcinoma cell line THC8307 / L-OHP

作　　者：李元杰[1,2] 张蕾[1] 隋御[1] 徐方[1,2] 

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750004 [2]宁夏医科大学生育力保持教育部重点实验室,宁夏银川750004

出　　处：《基础医学与临床》2014年第12期1623-1628,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(31360251);宁夏自然科学基金(NZ13079)

摘　　要：目的探讨沉默REV3和MMS2逆转结肠癌耐药细胞系THC8307/L-OHP细胞对奥沙利铂(L-OHP)的耐药性。方法 THC8307/L-OHP细胞转染含microRNA片段重组质粒后,用实时荧光定量PCR法(qRT-PCR)和免疫荧光法检测目的基因表达,选择低表达效率具有统计学意义的细胞作为实验组细胞。分别应用MTT法、罗丹明123实验、流式细胞术检测细胞对L-OHP药物敏感度和细胞凋亡。结果与对照组相比,L-OHP对实验组细胞的半数抑制浓度(IC50)及耐药指数(RI)减低(P<0.05),实验组细胞经L-OHP处理后凋亡率显著增高(P<0.05),实验组细胞MMS2和REV3蛋白表达水平降低(P<0.05)。结论下调REV3和MMS2在逆转人结肠癌细胞对L-OHP的耐药性和促进肿瘤细胞的凋亡上有一定作用。Objective To explore the effect of silencing REV3 and MMS2 gene in reversing oxaliplatin （L-OHP） resistance of human colon carcinoma THC8307/L-OHP cell line. Methods The real-time fluorescent quantitative PCR （qRT-PCR） was used to detect MMS2 expression in L-OHP-resistant THC8307/L-OHP cells and THC8307/L-OHP cells transfected with MMS2/REV3-RNAi plasmid recto or empty plasmid vector. The difference in MMS2 and REV3 expression was compared. MrlT assay, Rhodamine 123 assay and Flow-cytometric analysis were used to detect transfection of THC8307/L-OHP cells to L-OHP drug sensitivity, apoptosis on treatment with L-OHP. The protein expression of MMS2 and REV3 in THC8307/L-OHP cells was detected by immunofluoreseence. Results Compared to the control group, IC50 and resistance index（RI） of L-OHP in experimental group cells was lower （ P 〈 0.05 ） , apoptosis on treatment with L-OHP increased （ P 〈 0.05 ）. Furthermore, the protein expression of MMS2 and REV3 was downregulated in experimental group cells （ P 〈 0.05 ）. Conclu- sions Silencing REV3 and MMS2 gene from Oxaliplatin-resistant human colon cancer cells enhances the drug- sensitivity to L-OHP and induces apoptosis.

关 键 词：RNA干扰 MMS2 REV3 THC8307/L-OHP细胞 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]