miR-634 在肝癌中的表达及对肝癌细胞生物学行为的影响

目的:检测microRNA-634(miR-634)在肝癌中的表达水平及其对肝癌细胞常见生物学行为的调控作用。方法:采用实时荧光定量PCR(RT鄄qPCR)法检测肝癌细胞系(HepG2、SMMC7721、Bel7402、Bel7404、SNU739)、69 例肝癌组织及匹配癌旁组织中miR-634 的相对定量,分析miR-634 表达与肝癌患者性别、年龄、肿瘤直径、分化程度、Child-Pugh 分级、BCLC 分期、门静脉癌栓及肝外转移的关系,同时构建miR-634 的真核表达载体并转染肝癌细胞系,采用活细胞计数试剂盒CCK-8、流式细胞仪Annexin V/ PI 双染法和Transwell 侵袭实验检测转染miR鄄634 对细胞增殖、凋亡和侵袭能力的影响。结果:与正常人肝细胞系L-02 相比,肝癌细胞的miR-634 水平均降低(P <0.05),表达量依次为HepG2 >SNU739 >Bel7402 > Bel7404 >SMMC7721;69 例肝癌组织的miR鄄634 水平为(0.253±0.019),低于匹配癌旁组织(P<0.05),且与肿瘤直径、分化程度、BCLC分期、门静脉癌栓及肝外转移均有关(P<0.05)。过表达组转染24 ~96 h 后的miR鄄634 水平持续升高,与对照组和空转染组的差异有统计学意义(P<0.05);与对照组和空转染组相比,转染组的增殖抑制率、凋亡率均升高,但穿膜细胞数降低,差异有统计学意义(P<0.05)。结论:miR-634 在肝癌组织和细胞中均为低表达,且与临床病理参数有关,上调其水平可抑制肝癌细胞增殖及侵袭并诱导凋亡,对于肝癌防治有重要借鉴价值。

Expression of miR-634 in hepatocellular carcinoma and its effect on biological behavior of Hepatocellular carcinoma cells

Objective:To detect the expression level of microRNA-634 (miR-634) in hepatocellular carcinoma (HCC) and its regulatory effect on the common biological behavior of hepatocellular carcinoma cells.Methods: Real-time fluorescence quantitative PCR (RT qPCR) method was used to detect HCC cell lines (HepG2, SMMC7721, BEL7402, bel7404, SNU739), 69 cases of HCC tissues and matching relative quantification of miR-634 paracancerous tissues and analysis of relationship between miR-634 expression and HCC patients gender, age, tumor size, degree of differentiation, child Pugh classification, BCLC staging, portal vein tumor thrombus and liver metastasis, while building a miR-634 eukaryotic expression vector and transfected into hepatocellular carcinoma cell lines, using live cell counting kit-8 CCK-8, flow cytometric annexin V/ PI double staining and Transwell experiment to detect the transfection miR-634 on cell proliferation, apoptosis and invasion effects.Results: Compared with the normal human liver cell line L-02 and hepatocellular carcinoma cells miR-634 were decreased (P <0.05), the expression followed by HepG2 >SNU739 >Bel7402 > Bel7404>SMMC7721;69 cases of hepatocellular carcinoma (HCC) of miR-634 level (0.253±0.019) and lower than that of the matched paracancerous tissues (P<0.05), and related with the tumor size, degree of differentiation, BCLC stage, portal vein tumor thrombus and liver metastasis (P<0.05).Over expression in the transfected group 24-96 h after miR-634 level continues to rise, control group and blank vector transfected group differences were statistically significant (P<0.05);and the control transfection group and blank group compared to transfection proliferation inhibition rate, apoptosis rate was increased, but wear the number of cell membrane decreased, the difference was statistically significant (P<0.05).Conclusion: miR-634 in hepatocellular carcinoma tissues and cells were low expression and related to clinical pathological parameters, raised its level can inhibit the proliferation and invasion of hepatocellular carcinoma cells and induce apoptosis, for the prevention and treatment of liver cancer has an important reference value.