miR-133b通过靶向调控EGFR影响食管鳞癌的侵袭、转移

目的 探讨miR-133b在食管鳞癌侵袭、转移过程中的作用及机制.方法 使用生物信息学工具预测miR-133b的靶基因;双荧光素酶报告基因实验验证miR-133b对靶基因表皮生长因子受体(EGFR)的直接靶向调控作用;细胞增殖实验、划痕实验和Transwell实验检测miR-133b的不同表达对食管鳞癌细胞增殖、迁移和侵袭能力的影响;qRT-PCR验证miR-133b与EGFR在食管鳞癌组织中的表达,并分析miR-133b与食管鳞癌临床病理特征之间的关系.结果 生物信息学工具预测EGFR为miR-133b的靶基因,双荧光素酶报告基因实验证实 miR-133b可与 EGFR 3'UTR片段结合;体外实验通过上调食管鳞癌细胞中miR-133b的表达使细胞的增殖受阻(ECA109:P＜0. 05,KYSE150:P＜0. 01);细胞的迁移(ECA109:P＜0. 01,KYSE150:P＜0. 01)和侵袭能力(EC5A109:P＜0. 01,KYSE150:P＜0. 01)显著减弱.在食管鳞癌组织中 miR-133b与 EGFR的表达呈负相关性( P＜0. 01),且miR-133b的表达与食管鳞癌的TNM分期、淋巴结转移显著相关 (P均＜0. 05).结论 miR-133b可能通过下调EGFR的表达,抑制食管鳞癌的增殖、侵袭和转移,在食管鳞癌的演进中起负调控作用.

miR-133b regulates esophageal squamous cell carcinoma invasion and metastasis by targeting EGFR

Objective To investigate the role and mechanism of miR-133b in cell invasion and metastasis of esoph-ageal squamous cell carcinoma( ESCC) . Methods Bioinformatics tools were used to predict the potential target gene of miR-133b; dual luciferase reporter gene assay was conducted to verify the prediction. Cell proliferation, wound healing and transwell assay were performed to investigate the effects of miR-133b on cell proliferation, mi-gration and invasion in esophageal squamous cell carcinoma. qRT-PCR was conducted in a cohort of 33 pairs of ES-CC tissues and correspondingly adjacent tissues, then the relationship between the miR-133b and clinical pathologi-cal features weer analysed. Results Bioinformatics tools predicted that EGFR was the potential target gene of miR-133b, dual luciferase reporter gene assay verified that miR-133b could bind to a specific sequence of the 3′-UTR of the mRNA of EGFR. Overexpression of miR-133b in ESCC cell could inhibit proliferation(ECA109: P ＜0. 05, KYSE150: P＜0. 01), migration(ECA109: P＜0. 01, KYSE150: P＜0. 01) and invasion(ECA109:P＜0. 01, KYSE150: P＜0. 01) in vitro. The results demonstrated that EGFR, the target of miR-133b in ESCC, which up-regulated in ESCC tissues was negatively related to miR-133b(P＜0. 01), and miR-133b was frequently downregu-lated in esophageal squamous cell carcinoma tissues and its decrement correlated with TNM stage of ESCC and lym-phatic metastasis (P both＜0. 05). Conclusion MiR-133b may inhibit esophageal carcinoma proliferation, inva-sion and metastasis by targeting EGFR, it plays a negative role in the progression of ESCC.