MicroRNA-338-3p通过下调SMO表达抑制结直肠癌细胞的侵袭与迁移  

MicroRNA-338-3p suppresses invasion and migration of colorectal cancer cells by targeting SMO

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：《中国现代医学杂志》2013年第18期25-30,共6页China Journal of Modern Medicine

摘　　要：目的探讨microRNA-338-3p(miR-338-3p)对结直肠癌细胞侵袭与迁移能力的影响及其调控机制。方法脂质体分别转染miR-338-3p前体(pre-miR-338-3p)和miR-338-3p特异性抑制剂(miR-338-3p-inhibitor)至人结直肠癌SW620及SW480细胞系中,应用实时荧光定量PCR(Real-time RT-PCR)检测细胞中miR-338-3p表达状况,运用半定量RT-PCR及Westernblot分析Smoothened(SMO)mRNA及蛋白表达状况,Transwell侵袭、迁移实验检测转染前后细胞体外侵袭迁移能力;在经anti-SMO-siRNA预先处理的SW480细胞中转染miR-338-3p-inhibitor,检测转染细胞中SMO蛋白的表达变化以及侵袭能力的改变。结果通过生物信息学网站预测SMO可能是miR-338-3p的作用靶基因;SW620细胞中miR-338-3p表达水平较SW480细胞减低,而SMO蛋白增高。转染pre-miR-338-3p至SW620细胞,miR-338-3p表达上调,SMO蛋白表达降低,且肿瘤细胞的侵袭及迁移能力下降;而转染miR-338-3p-inhibitor至SW480细胞后,miR-338-3p表达下降,SMO蛋白表达上升,肿瘤细胞侵袭、迁移能力增强,且此种增强效应可被an-ti-SMO-siRNA所部分逆转,说明miR-338-3p确实是通过抑制SMO而发挥抗癌作用。结论 MiR-338-3p通过下调SMO表达而抑制结直肠癌细胞侵袭与迁移。To explore the effects of microRNA-338-3p（miR-338-3p） on the invasion and migration ability of colorectal cancer（CRC） cells and the mechanisms.【Methods】MiR-338-3p precursor（pre-miR-338-3p） or miR-338-3p specific inhibitor（miR-338-3p-inhibitor） were transfected into human colorectal cancer cell lines SW620 and SW480,respectively.Real-time fluorescence quantitative PCR（Real-time RT-PCR） was used to detect miR-338-3p expression.SMO mRNA and protein were analyzed by semi-quantitative RT-PCR and Western-blot,respectively.Furthermore,the vitro cell invasion and migration ability were detected by Transwell invasion and migration assay.Moreover,miR-338-3p-inhibitor was transfected into anti-SMO-siRNA-pretreated SW480 cells,and then the SMO protein and invasive and migration ability of the cells were detected.【Results】SMO was predicted to be the target gene of miR-338-3p by bio-information websites.MiR-338-3p expression in SW620 cells was lower than that in SW480,but SMO protein was increased.After the transfection of pre-miR-338-3p into SW620 cells,miR-338-3p expression was up-regulated,SMO protein expression was down-regulated,and the invasion and migration ability of the cells were decreased;however,miR-338-3p expression was decreased and SMO protein was increased after transfection of miR-338-3p-inhibitor into SW480 cells.Moreover,the invasion and migration ability of SW480 cells were also reinforced and this effect could be partially reversed by anti-SMO-siRNA,suggesting that the regulative effect of miR-338-3p was indeed mediated by SMO.【Conclusions】MiR-338-3p suppress invasion and migration ability of CRC by targeting SMO.

关 键 词：结直肠癌 MicroRNA-338-3p SMO SHH信号通路 侵袭 迁移 

分 类 号：R735.35[医药卫生—肿瘤]