【摘要】 目的探讨Notch信号通路受体分子在肝细胞癌(HCC)中的表达及临床意义,筛选可能调控Notch2基因的microRNAs。方法应用RT-qPCR检测38例HCC和癌旁肝组织Notch受体mRNA的表达,寻找关键的Notch信号受体;应用组织芯片免疫组织化学法检测106例HCC和癌旁肝组织Notch2蛋白的表达,分析与临床病理特征的相关性。Western-blot检测4例HCC和癌旁肝组织Notch2蛋白的表达。通过生物信息数据库预测Notch2可能相关的microRNAs,应用RT-qPCR检测46例HCC和正常肝组织中microRNAs和Notch2的表达,筛选出差异表达的microRNAs。Pearson′s相关分析HCC组织中Notch2和差异microRNAs的关系。结果 Notch2 mRNA在HCC组织中的相对表达量明显高于癌旁肝组织;Notch1、Notch3和Notch4 mRNA在2种组织中的表达差别无统计学意义。HCC组织的Notch2蛋白表达水平高于癌旁肝组织,Notch2蛋白在复发、肝外转移及B-D期的HCC患者中高表达。HCC组织的miR-146a表达量明显低于正常肝组织,而miR-16、miR-30b、miR-181a和miR-29b在2种组织中的表达差别无统计学意义。HCC组织中Notch2和miR-146a表达呈明显负相关。结论 Notch2可能是HCC组织中关键的Notch信号通路受体分子;Notch2蛋白表达可作为预测HCC复发、转移的生物标志物;Notch2可能是miR-146a调控的靶基因。更多还原

【Abstract】 Objective To investigate the expression and clinical significance of Notch signal pathway receptor molecules in hepatocellular carcinoma(HCC), and to screen the related microRNAs. Methods Quantitative real-time PCR(RT-qPCR) was used to detect the expression of Notch receptors in 38 cases of HCC and adjacent liver tissues to find the key Notch signal receptors. The expression of Notch2 protein in 106 cases of HCC and adjacent liver tissues was detected by tissue microarray immunohistochemistry, and analyzed the clinicopathological features. Western-blot was used to detect the expression of Notch2 in 4 cases of human HCC and adjacent liver tissues. Expression of microRNAs and Notch2 in 46 cases of HCC and normal liver tissues were examined by RT-qPCR. The microRNAs related to Notch2 were predicted using bioinformatics. Differential expression of microRNAs was screened out in 46 cases of HCC. Results The level of Notch2 mRNA in HCC was significantly higher than that in adjacent liver tissues, while the expressions of Notch1, Notch3, and Notch4 were not significantly different. Notch2 protein expression in HCC was significantly higher than adjacent liver tissues. Notch2 protein was highly expressed in patients with recurring, extrahepatic metastasis and B-D phase. The expression of miR-146 a in HCC was significantly lower than that in normal liver tissues, while the expression of miR-16, miR-30 b, miR-181, and miR-29 b in HCC was not significantly different compared to normal liver tissues. Pearson′s correlation analysis showed that the expression of Notch2 had a significantly negative correlation with miR-146 a in HCC. Conclusion Notch2 is the key receptor molecule of the Notch signaling pathway in HCC and the high expression of Notch2 protein can be used as a biomarker to predict the recurrence and metastasis of HCC. Notch2 may be a target gene of miR-146 a.更多还原