【作者】 徐洁；

【导师】 吴福生；

【作者基本信息】 浙江大学 ， 肿瘤学， 2011， 硕士

【摘要】 目的：检测microRNA-122 (miR-122)在肝细胞肝癌组织及肝癌细胞株的表达,分析miR-122的表达量同肝癌临床病理因素之间的相关性,并探讨(?)miR-122对肝癌细胞生物学行为的影响,为肝细胞肝癌的靶向治疗提供实验依据。方法：应用实时荧光定量PCR技术检测30例肝癌组织及其配对癌旁组织、8种肝癌细胞株中miR-122的表达情况；分析miR-122的表达量同肝癌临床病理特征之间的相关性；肝癌细胞株HepG2转染miR-122 Agomir使其过表达miR-122,通过细胞增殖检测试剂盒CCK-8检测过表达miR-122对HepG2细胞增殖能力的影响；流式细胞仪检测过表达miR-122对HepG2细胞凋亡的影响。所有资料采用SPSS 16.0 for Windows软件处理,计量数据均以均数±标准差(means±SD)表示,采用t检验、方差分析,P<0.05被认为有统计学意义。结果：miR-122在肝癌组织、肝癌细胞株中(除Huh-7细胞外)低表达,肝癌组织中:miR-122表达量低于其癌旁组织者有27例,表达量相仿或高于其癌旁组织者3例,差异具有统计学意义(P<0.01)。在8种肝癌细胞株中,除Huh-7细胞中miR-122表达量高于Chang liver正常肝细胞外,其它7种肝癌细胞中miR-122表达量均低于Chang liver正常肝细胞(P<0.01),且高转移潜能的肝癌细胞株MHCC-97H.HCCLM3中miR-122的表达量低于低转移潜能的肝癌细胞MHCC-97L(P<0.01),差异具有统计学意义。miR-122的表达量同肝癌临床病理因素之间的相关性分析结果显示miR-122的表达量同肝癌分化程度呈正相关(P<0.01)。CCK-8结果显示过表达miR-122抑制了HepG2细胞的增殖(P<0.01)。流式细胞仪检测结果显示过表达miR-122促进了HepG2细胞的凋亡(P<0.01)。结论：(1)miR-122在肝癌中低表达,其表达量与肝癌分化程度正相关。(2)过表达miR-122具有抑制HepG2肝癌细胞的增殖、促进细胞凋亡的作用。更多还原

【Abstract】 ’Objective:To investigate the expression level of microRNA-122 (miR-122) in hepatocellular carcinoma(HCC) tissues and cell lines, analysis the potential relationship between miR-122 expression and clinicopathologic parameters in HCC patients, explore the biological functions of miR-122 and provide a basis for the targeting treatment of HCC.Methods:The expression level of miR-122 in hepatocellular carcinoma tissues and cell lines were quantified by using TaqMan MicroRNA Assays-based real-time RT-PCR. The correlations of clinicalpathologic parameters with the miR-122 expression level in cancerous tissues were analyzed by using independent-Samples T Test respectively. MiR-122 Agomir was transfected into HepG2 hepatocellular carcinoma cell to overexpress miR-122. The effect of over-expression miR-122 on HepG2 cell proliferation and apoptosis were evaluated by using CCK-8 kit and flow cytometer respectively. All the data were expressed as means±SD and analyzed by using SPSS 16.0 for Windows. Student t and ANOVA were used to compare the differences between corresponding groups. P<0.05 was regarded as statistically significant.Results:The expression level of miR-122 in HCC tissues was significantly decreased than the expression in paracancerou tissues (P<0.01). Compared to Chang liver cell, the expression level of miR-122 was significantly decreased in all HCC cell lines except for Huh-7 (P<0.01). The highly metastatic variant, MHCC97-H and HCCLM3 had a lower miR-122 expression than the low metastatic potential variant, MHCC97-L (P< 0.01). The expression level of miR-122 was significantly correlated with the HCC differentiation grade (P<0.01). However, no significant correlation was observed between other clinicalpathologic parameters and the expression level of miR-122 in HCC patients respectively (P>0.05). In vitro experiment, over-expression of miR-122 significantly inhibited the proliferation(P<0.01), and prbmoted the apoptosis of HepG2 cell (P<0.01), compared to the HepG2-negative group and the HepG2-control group cells.Conclusions:The expression level of miR-122 was significantly down-regulated in HCC tissues and cell lines, and its down-regulation was correlated with poor differentiation of hepatocellular carcinoma. Over expression of miR-122 could inhibit HepG2 cell growth and promote the cell apoptosis.更多还原