原发性肝细胞癌组织miR-221表达变化及其对肿瘤侵袭和转移的影响

Expression of miR-221 in primary hepatocellular carcinoma and its effect on invasion and metastasis of hepatocellular carcinoma

目的探讨原发性肝细胞癌(HCC)组织微小RNA-221(miR-221)表达变化及其对肿瘤侵袭和转移的影响。方法采用RT-qPCR法检测74例份HCC组织及其配对的癌旁正常组织miR-221表达。以miR-221表达的均数为截断值,将HCC组织分为miR-221高表达与miR-221低表达,分析不同miR-221表达与患者临床病理特征的关系。体外传代培养高侵袭性HCC细胞MHCC97H、低侵袭性HCC细胞MHCC97L及正常肝细胞L02,采用RT-qPCR法检测各细胞miR-221表达,选择miR-221相对表达量最高的细胞系进行后续实验。将上述miR-221相对表达量最高的细胞随机分为观察组和对照组,分别转染miR-221inhibitor和NCinhibitor。转染24h,收集细胞,采用细胞-基质黏附实验分别检测Fibronectin或Matrigel胶包被时细胞黏附能力;采用Transwell侵袭和迁移实验检测细胞侵袭和迁移能力。结果HCC组织miR-221相对表达量明显高于癌旁正常组织(P<0.05)。74例份HCC组织中,miR-221高表达39例份、miR-221低表达35例份。HCC组织miR-221高表达与Edmondson-Steiner分级、微血管侵犯和临床分期有关(P均<0.05),与性别、年龄、肿瘤直径、肿瘤数目、肝硬化、HBV感染无关(P均>0.05)。高侵袭性HCC细胞MHCC97H和低侵袭性HCC细胞MHCC97L miR-221相对表达量明显高于正常肝细胞L02,且高侵袭性HCC细胞MHCC97HmiR-221相对表达量明显高于低侵袭性HCC细胞MHCC97L(P均<0.05),故以高侵袭性HCC细胞MHCC97H进行后续实验。观察组无论是Fibronectin包被时还是Matrigel胶包被时细胞黏附能力均明显低于对照组(P均<0.05)。观察组细胞侵袭和迁移能力均明显低于对照组(P均<0.05)。结论HCC组织miR-221表达明显升高,其高表达可促进肿瘤侵袭和转移;下调miR-221表达能够抑制HCC细胞的黏附、侵袭和迁移能力。

ObjectiveTo investigate the expression of miR-221 in the primary hepatocellular carcinoma (HCC) and its relationship with tumor progression. Methods The expression of miR-221 was detected by qRT-PCR in 74 HCC tissues and their matched normal adjacent tissues. The mean expression of miR-221 in the HCC tissues was used as the cut-off value. The patients were divided into high expression of miR-221 and low expression of miR-221. The relationship between the expression of miR-221 and the clinicopathological characteristics of patients was analyzed.High invasive HCC cells MHCC97H, low invasive HCC cells MHCC97L, and normal hepatocytes L02 were cultured in vitro. The expression of miR-221 was detected by qRT-PCR. The cell lines with the highest relative expression of miR-211 were selected for the subsequent experiments. The cells with the highest relative expression of miR-211 were randomly divided into the observation group and control group, and transfected with miR-221 inhibitor and NC-inhibitor for 24 h, respectively. Cell-matrix adhesion assay was used to detect the cell adhesion ability when they were coated by Fibronectin or Matrigel. Transwell invasion and migration assays were used to detect the cell invasion and migration. Results The relative expression of miR-221 in the HCC tissues was significantly higher than that in the adjacent normal tissues (P<0.05). Among 74 cases of HCC tissues, 39 cases had high expression of miR-221 and 35 cases had low expression of miR-221. The high expression of miR-221 in the HCC tissues was related to Edmondson-Steiner classification, microvascular invasion and clinical stage (all P<0.05), but not to sex, age, tumor diameter, number of tumors, cirrhosis, or HBV infection (all P>0.05). Compared with the normal hepatocyte line L02, the relative expression of miR-221 in the MHCC97H cells and MHCC97L cells was significantly higher;the relative expression of miR-221 in MHCC97H cells was significantly higher than that in MHCC97L cells (all P<0.05). Therefore, MHCC97H cells were used for the subsequent experiments. The cell adhesion ability of the observation group was significantly lower than that of the control group in both Fibronectin and Matrigel gel coatings (all P< 0.05 ). The cell invasion and migration abilities of the observation group were significantly lower than those of the control group (both P<0.05). Conclusion The expression of miR-221 in the HCC tissues increases significantly, and its high expression can promote the invasion and metastasis of tumors;down-regulation of miR-211 expression can inhibit the adhesion, invasion, and migration of HCC cells.