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.
出处
《基础医学与临床》
CSCD
北大核心
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)
