mir-338-3p对结直肠癌细胞侵袭迁移能力及SMO蛋白表达的影响  

Effects of microRNA-338-3p on invasion and migration and SMO protein expression in human colorectal cancer cells

作　　者：区文弢[1] 孙凯[1] 吴承堂[1] 雷尚通[1] 张晓槟[1] 

机构地区：[1]南方医科大学南方医院普外科,广东广州510515

出　　处：《广州医学院学报》2013年第3期1-5,共5页Academic Journal of Guangzhou Medical College

基　　金：国家自然科学基金青年科学基金项目(81101896)

摘　　要：目的:探讨微小RNA-338-3p(miR-338-3p)对人结直肠癌细胞侵袭迁移能力及Smoothened(SMO)蛋白表达的影响。方法:应用实时荧光定量PCR(Real-time PCR)检测四种人结肠癌细胞系Lovo、HT-29、HCT116、SW620中miR-338-3p表达状况,并以人脐静脉内皮细胞(HUVEC)作为对照。采取脂质体转染miR-338-3p前体(pre-miR-338-3p)至人结直肠癌细胞系SW620,观察细胞转染前后miR-338-3p的表达变化,Transwell侵袭实验及划痕实验检测转染前后细胞侵袭及迁移能力的改变;以Western-blotting和RT-PCR分析SMO蛋白及mRNA表达状况。结果:检测四种结直肠癌细胞系中miR-338-3p表达水平均较正常对照细胞显著降低,差异具有统计学意义(P<0.01);SW620细胞转染pre-miR-338-3p后,miR-338-3p表达水平升高,SMO蛋白表达下降,同时肿瘤细胞侵袭、迁移能力明显增强,与对照组相比差异具有统计学意义(P<0.01);但SMO mRNA表达水平在转染前后差异无统计学意义(P>0.05)。结论:MiR-338-3p可通过抑制结直肠癌细胞系中SMO蛋白表达从而抑制肿瘤细胞侵袭转移。Objective： To investigate the effects of microRNA-338-3p（miR-338-3p） on cell invasion and migration and Smoothened protein（SMO） expression in human colorectal cancer（CRC） cells. Methods： Real-time polymerase chain reaction（ RT-PCR）was employed to detect the miR-338-3p expression in Lovo, HT-29, HCT116 and SW620, the four types of human CRC cell lines, and in human umbilical vein endothelial cells （HUVEC） , the controls. Human CRC cell line,SW620,was transfected with a miR-338-3p precursor（pre-miR-338-3p） by using liposomes. This allowed determination of the changes in miR-338-3p expression, and CRC cell invasion and migration by Transwell and scratch assays following transfection. Western-blotting and semi-quantitative RT-PCR were adopted to analyze Smoothened（SMO） protein and mRNA expression. Results：Compared with control cells, all human CRC cell lines yielded a significantly attenuated expression of miR-338-3p（ all P 〈 0.01 ）. Transfection of 5W620 cells with pre-miR-338-3p, when compared with control cells ,was associated with augmented miR-338- 3p expression,attenuated SMO protein expression and improved capacity of invasion and migration（ all P 〈 0. 01 ） , but not the expression of SMO mRNA （P 〉 0.05 ）. Conclusion： MiR-338-3p suppresses CRC invasion and migration via inhibiting SMO protein expression.

关 键 词：结直肠癌 微小RNA Smoothened基因 

分 类 号：R574.62[医药卫生—消化系统]