miRNA-490-3p调控Smad2/TGF-β影响结直肠癌SW480细胞的奥沙利铂敏感性  

Effect of miRNA-490-3p regulating Smad2/TGF-βon sensitivity of colorectal cancer SW480 cells to oxaliplatin

作　　者：李志发[1] 吴小兵[1] 罗超元[1] 陈戎[1] LI Zhifa;WU Xiaobing;LUO Chaoyuan;CHEN Rong(Department of Gastrointestinal Surgery,the Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150,Guangdong,China)

机构地区：[1]广州医科大学附属第三医院胃肠外科,广东广州510150

出　　处：《中国肿瘤生物治疗杂志》2021年第5期489-494,共6页Chinese Journal of Cancer Biotherapy

基　　金：广东省医学科技技术研究基金资助项目(No.A2016500);广东省自筹经费类科技计划资助项目(No.2017ZC0250)。

摘　　要：目的:探讨miRNA-490-3p调控Smad2/TGF-β对结直肠癌奥沙利铂化疗敏感性的影响。方法:选取100例结直肠癌(colorectal cancer,CRC)根治性手术后奥沙利铂(oxaliplatin,OXA)同种联合化疗患者作为研究对象,根据化疗情况分为耐药组(n=40)和非耐药组(n=60),用q PCR检测两组外周血miRNA-490-3p水平;选取人CRC细胞株SW480和CRC奥沙利铂(OXA)耐药型细胞株OXA-SW480进行研究,先用q PCR检测两种细胞株中miRNA-490-3p表达水平;后将miRNA-490-3p过表达载体转染OXA-SW480(过表达组),并设立空载体组(空载体转染OXA-SW480)和空白对照组(OXA-SW480未经任何处理)。用CCK-8检测各组细胞增殖能力,同时用不同浓度OXA处理各组细胞,计算其半数抑制浓度(IC50);用Annexin-V-FITC/PI染色流式细胞术检测各组细胞凋亡率;用WB检测各组Smad2和TGF-β蛋白表达水平。结果:在CRC患者中,耐药组外周血miR-490-3p水平显著低于非耐药组,在CRC细胞株中,OXA-SW480耐药株细胞miR-490-3p水平显著低于正常株SW480细胞(P<0.05)。与空载体组和空白对照组相比,过表达组miR-490-3p水平显著降低(均P<0.05),增殖抑制率显著升高(均P<0.05),细胞凋亡率显著升高(均P<0.01),Smad2蛋白水平显著降低(均P<0.05),TGF-β蛋白水平显著升高(均P<0.05)。经OXA处理后,过表达组的IC50显著低于空载体组和空白对照组(均P<0.05)。经Pearson相关法分析,miR-490-3p与Smad2的表达呈负相关(r=–0.943,P<0.01),miR-490-3p与TGF-β的表达呈正相关(r=0.961,P<0.01)。结论:过表达miRNA-490-3p可增加CRC SW480细胞的OXA敏感性,此作用Smad2/TGF-β信号通路有关。Objective:To investigate the effect of miRNA-490-3 p regulating Smad2/TGF-βon chemo-sensitivity of colorectal cancer to oxaliplatin.Methods:One hundred patients with colorectal cancer(CRC)who received oxaliplatin(OXA)chemotherapy after radical operation were selected as research subjects and divided into drug resistant group(n=40)and non-resistant group(n=60)according to the chemotherapy outcome.The level of miRNA-490-3 p in peripheral blood of two groups of patients was detected by qPCR.Human CRC cell line SW480 and oxaliplatin resistant CRC cell line OXA-SW480 were selected for the study.The expression level of miRNA-490-3 p in the two cell lines was detected by qPCR,and then the miRNA-490-3 p over-expression vector was transfected into OXA-SW480 cells(over-expression group),in the meanwhile,the empty vector group(OXA-SW480 cells transfected with empty vector)and blank control group(OXA-SW480 cells without any treatment)were set up.CCK-8 was used to detect cell proliferation ability of each group,and different concentrations of OXA were used to treat cells of each group to calculate the half inhibitory concentration(IC50);Annexin-V-FITC/PI staining was used to detect cell apoptosis rate of each group;WB assay was used to detect the expression levels of Smad2 and TGF-β.Results:In CRC patients,the level of miR-490-3 p in the peripheral blood of the drug-resistant group was significantly lower than that of the non-resistant group.In the CRC cells,the level of miR-490-3 p in the OXA-SW480 cells was significantly lower than that of the normal SW480 cells(P<0.05).Compared with the empty vector group and the blank control group,the level of miR-490-3 p in the over-expression group was significantly reduced(all P<0.05),the proliferation inhibition rate and the apoptosis rate were significantly increased(all P<0.05),Smad2 protein level was significantly reduced(all P<0.05),and TGF-βprotein level was significantly increased(all P<0.05).After OXA treatment,the IC50 of the over-expression group was significantly lower t

关 键 词：miRNA-490-3p 结直肠癌 奥沙利铂 耐药 机制 

分 类 号：R735.3[医药卫生—肿瘤] R730.53[医药卫生—临床医学]