miR-206通过靶定Arpc3抑制食管癌细胞的迁移和侵袭

MiR-206 Inhibits Migration and Invasion of Esophageal Cancer Cells by Targeting Arpc3

目的探讨miR-206在食管癌组织和细胞中的表达,及其对食管癌细胞迁移和侵袭能力的影响。方法 qRT-PCR检测miR-206在食管癌组织和细胞系中的表达水平;慢病毒感染获得稳定过表达miR-206的TE-1细胞,划痕实验和Transwell实验检测细胞的迁移及侵袭能力;生物信息学预测miR-206与Arpc3可能的结合位点,并通过双荧光素酶报告实验验证其靶向调控关系;通过siRNA技术降低TE-1细胞中Arpc3的表达,并检测其对细胞迁移和侵袭能力的影响。结果与癌旁组织比较,食管癌组织中miR-206表达水平明显下调(P<0.01);与正常人食管上皮细胞系比较,食管癌细胞系(TE-1、KYSE-30、KYSE-70和Eca-109)miR-206表达水平明显下调(均P<0.01);过表达miR-206能显著抑制TE-1细胞的迁移和侵袭(均P<0.01);Arpc3是miR-206的直接靶基因,过表达miR-206后TE-1细胞中Arpc3表达降低(P<0.01);抑制Arpc3表达能抑制TE-1细胞的迁移和侵袭(均P<0.05)。结论 miR-206通过靶向调控Arpc3而抑制食管癌细胞的迁移和侵袭,在食管癌中发挥抑癌基因的作用。

Objective To investigate the expression of miR-206 in esophageal cancer tissues and cell lines,and determine its effects on the migration and invasion of esophageal cancer cells. Methods The expression of miR-206 was detected in esophageal cancer tissues and cell lines by quantitative real-time PCR(qRT-PCR).A stable esophageal cancer cell line(TE-1)overexpressing the miR-206 was constructed,and the cell migration and invasion were analyzed by wound healing assay and Transwell chamber assay.The potential binding site of miR-206 on the Arpc3 was predicted by using on-line bioinformative databases,and the result was verified by double luciferase assay.The migration and invasive ability of TE-1 cells were detected after down-regulating the Arpc3 by siRNA technology. Results The expression level of miR-206 was significantly lower in esophageal cancer tissues than in adjacent normal tissues(P<0.01).Similarly,miR-206 expression in the four esophageal cancer cell lines(TE-1,KYSE-30,KYSE-70 and Eca-109)was significantly lower than that in the normal esophageal epithelial cell lines HEEC(P<0.01).Overexpression of miR-206 in TE-1 cells reduced cell migration and invasion(P<0.01).Arpc3 is a target gene of miR-206 and overexpression of miR-206 significantly decreased expression of Arpc3 in TE-1 cells(P<0.01).Downregulation of Arpc3 significantly reduced cell migration and invasion(P<0.05). Conclusion miR-206 inhibits migration and invasion of esophageal cancer cells by targeting Arpc3,and may serve as a tumor suppressor.