目的：探讨hsamiR132（miR132）对人食管癌细胞增殖和侵袭的影响及其可能的机制。方法：构建pCDNA3.1(+)miR132表达载体并转染高转移潜能的人食管癌细胞KYSE150，G418筛选稳定过表达miR132的KYSE150细胞。软琼脂克隆形成实验和Transwell小室实验检测KYSE150细胞的增殖和侵袭，Realtime PCR和Western blotting检测miR132和叉头框蛋白A1基因（forkhead box protens A1，FOXA1）在KYSE150细胞中的表达，双荧光素酶报告基因分析和Western blotting检测过表达miR132对FOXA1的调控作用。结果：成功构建pCDNA3.1(+)miR132质粒，获得稳定过表达miR132的KYSE150细胞。亲本KYSE150细胞中miR132低表达，而FOXA1蛋白高表达。与转染空载体和未转染KYSE150细胞相比，转染pCDNA3.1(+)miR132不影响KYSE150细胞的增殖（13.9±0.33 vs 15.4±0.11、17.1±0.20，P>0.05），但细胞体积较小且表面比较光滑；其可显著降低KYSE150细胞的侵袭能力\[（55±1.6) vs (129.0±3.1)、(124.0±2.8)个细胞，P<0.01\]。miR132能够作用于FOXA1 3′UTR，从而抑制内源性FOXA1的表达。结论：食管癌KYSE150细胞低表达miR132，外源性过表达miR132可负向调控FOXA1的表达，从而抑制KYSE150细胞的侵袭能力。

Objective:To investigate the effect of hsamiR132 (miR132) on proliferation and invasion of human esophageal carcinoma cells, and further explore its possible mechanism. Methods: pCDNA3.1(+)miR132 plasmid was constructed, and KYSE150 cells stablely overexpressing miR132 were selected through G418 screening. Soft agar colony formation test and Transwell assay were performed to analyze the proliferation and invasion of KYSE150 cells. Expression of miR132 and forkhead box protens A1 (FOXA1) in KYSE150 cells were examined by realtime PCR and Western blotting. The regulatory effect of miR132 overexpression on FOXA1 was further studied by reporter gene assay and Western blotting. Results: pCDNA3.1(+)miR132 plasmid was successfully constructed, and KYSE150 cells stablely overexpressing miR132 was established. In parental KYSE150 cells, miR132 expression was low ,while FOXA1 expression was high. Compared to mock transfected KYSE150 cells and untransfected KYSE150 cells, pCDNA3.1(+)miR132 transfection did not affect the proliferation of KYSE150 cells (13.9±0.33 vs 15.4±0.11, 17.1±0.20, P>0.05), however tumor size reduced and tumor appeared smoother after pCDNA3.1(+)miR132 transfection. Furthermore, overexpression of miR132 significantly suppressed the invasion of KYSE150 cells（55±1.6 vs 129.0±3.1, 124.0±2.8, P＜001), and miR132 decreased the expression of endogenous FOXA1 by targeting FOXA1 3'UTR. Conclusion: Esophageal carcinoma KYSE150 cells express low level of miR132, and overexpression of miR132 can negatively regulate the expression of FOXA1 and suppressed the invasion of KYSE150 cells.

河北省卫生厅重点课题计划资助项目（No.20110068）