miR-30靶向调控Zeb2相关通路抑制胆囊癌细胞EMT进程

miR-30 inhibits EMT of gallbladder cancer cells via Zeb2-related pathways

目的 本研究旨在探讨小RNA-30(miR-30)在胆囊癌(GBC)上皮间充质转化(EMT)进程中的作用和潜在机制。方法 qRT-PCR用于检测胆囊上皮细胞和癌细胞系miR-30相对表达水平;过表达miR-30后采用MTT、平板克隆实验用于检测细胞增殖能力;细胞划痕实验和Transwell小室试验分别评估细胞迁移和侵袭能力;Western blot用于评估细胞EMT相关蛋白(E-cadherin、N-cadherin和Vimentin)和EMT相关转录因子(Snail、Slug和Zeb2)的表达水平。拯救实验用于探索miR-30与Zeb2的靶向调控关系。结果 miR-30在2个代表性GBC细胞系(GBC-SD和NOZ)中明显下调(P<0.05)。过表达miR-30会抑制GBC-SD细胞增殖、克隆形成、细胞迁移、细胞侵袭和EMT过程(P<0.05),而过表达Zeb2可以逆转miR-30导致的EMT抑制作用(P<0.05)。结论 miR-30通过靶向Zeb2减弱GBC细胞的EMT进展,表明miR-30是一种肿瘤抑制因子,可作为GBC的新型潜在分子治疗靶点。

Objective This study aimed to explore the role and potential mechanism of miR-30 in EMT of gallbladder cancer(GBC).Methods qRT-PCR was used to measure the relative expression level of miR-30 in gallbladder epithelial cells and cancer cell lines.After overexpression of miR-30,MTT and colony formation assays were used to assess cell proliferation.Wound healing and Transwell assays were used to evaluate cell migratory and invasive abilities,respectively.Western blot was used to evaluate the expression levels of EMT-related proteins(E-cadherin,N-cadherin and Vimentin) and EMT-related transcription factors(Snail,Slug and Zeb2).A rescue experiment was carried to explore the regulatory relationship between miR-30 and Zeb2.Results miR-30 was significantly downregulated in two representative GBC cell lines(GBC-SD and NOZ)(P<0.05).Overexpression of miR-30 inhibited GBC-SD cell proliferation,colony formation,migration,invasion and EMT(P<0.05).Overexpression of Zeb2 reverses the EMT inhibitory effect caused by miR-30(P<0.05).Conclusions miR-30 attenuates EMT of GBC cells by targeting Zeb2,which indicates that miR-30 is a tumor suppressor and can be used as a novel therapeutic target for GBC.