Yao Bowen, Xue Yumo, Liu Zhikui, Xu Meng, Tu Kangsheng, Wang Jun
Department of Hepatobiliary Surgery, First Affiliated Hospital, Medical College, Xi'an Jiaotong University, Xi'an 710061, China.
Department of Emergency, First Affiliated Hospital, Medical College, Xi'an Jiaotong University, Xi'an 710061, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Aug;32(8):1083-7.
Objective To observe the expression level of miR-141 in tumor tissues of human hepatocellular carcinoma (HCC) and determine the effect of miR-141 level on cell proliferation, invasion and migration of MHCC-97H cells by upregulation of miR-141. Methods We checked the miR-141 expression level in HCC by real-time quantitative PCR and analyzed the relationship between the expression level of miR-141 and clinical pathological indicators as well as survival rate. MHCC-97H cells were transiently transfected with miR-141 mimics which were artificially synthesized. The proliferation of MHCC-97H cells was detected by MTT assay. Transwell(TM) assay was performed to examine the invasion and migration of MHCC-97H cells. The expression of erythropoietin-producing hepatocellular receptor A2 (EphA2), which was the potential downstream target, was determined by Western blotting and immunohistochemistry. Results The expression level of miR-141 in HCC tissues was significantly lower than that in the adjacent normal tissues, and it was obviously associated with TNM stage, portal vein infiltration and Edmondson degree. Patients in the lower miR-141 group had a worse 3-year survival than those in higher miR-141 group. Overexpression of miR-141 in MHCC-97H cells significantly suppressed cell proliferation, invasion and migration, and inhibited the protein expression of EphA2. Correlation analysis showed that miR-141 level was negatively correlated with EphA2 expression level. Conclusion miR-141 is down-regulated in HCC tissues and it is negatively correlated with EphA2 expression. Its low expression is correlated with the malignant clinical pathological features. miR-141 overexpression down-regulates EphA2 expression and subsequently inhibits the proliferation, invasion and migration of HCC cells.
目的 观察miR-141在人肝细胞癌(HCC)肿瘤组织中的表达水平,并通过上调miR-141来确定其水平对MHCC-97H细胞增殖、侵袭和迁移的影响。方法 采用实时定量PCR检测HCC中miR-141的表达水平,并分析miR-141表达水平与临床病理指标及生存率之间的关系。用人工合成的miR-141模拟物对MHCC-97H细胞进行瞬时转染。采用MTT法检测MHCC-97H细胞的增殖情况。进行Transwell(TM)实验检测MHCC-97H细胞的侵袭和迁移能力。通过蛋白质印迹法和免疫组织化学法检测潜在下游靶点促红细胞生成素产生肝细胞受体A2(EphA2)的表达。结果 HCC组织中miR-141的表达水平明显低于癌旁正常组织,且与TNM分期、门静脉浸润及Edmondson分级明显相关。miR-141低表达组患者的3年生存率低于miR-141高表达组。miR-141在MHCC-97H细胞中的过表达显著抑制细胞增殖、侵袭和迁移,并抑制EphA2的蛋白表达。相关性分析显示,miR-141水平与EphA2表达水平呈负相关。结论 miR-141在HCC组织中表达下调,且与EphA2表达呈负相关。其低表达与临床恶性病理特征相关。miR-141过表达下调EphA2表达,进而抑制HCC细胞的增殖、侵袭和迁移。