CCN3对肝细胞肝癌侵袭的作用及相关机制的研究

发布时间：2018-04-09 23:18

本文选题：CCN3　切入点：肝细胞肝癌　出处：《南京医科大学》2015年硕士论文

【摘要】：[目的]CCN3(NOV,Nephroblastoma overexpressed gene,肾母细胞瘤过表达基因)是CCN家族的一个成员,参与多种细胞功能的发生和发展,包括细胞生长,分化,血管生成,粘附等。目前研究者们发现CCN3在多种恶性肿瘤中高表达,调控多种肿瘤的发生与发展,如骨肉瘤,食管癌等。然而CCN3在肝细胞肝癌中的相关研究尚未报道。在本研究中,我们对CCN3在肝癌组织及细胞中的表达量进行检测,并进行相应的功能实验,探讨相应机制。[方法]采用实时定量聚合酶链式反应(RT-PCR)实验和蛋白免疫印迹法(Western bolt)检测50对肝癌组织及其癌旁组织中CCN3的表达水平,统计分析CCN3的表达水平和临床肝癌患者病例资料的相关性。同时检测肝癌细胞株MHCC-97H,SMMC-7721,Hep G2,Huh7,MHCC-97L及正常人类肝细胞L02中CCN3的m RNA表达水平,构建CCN3过表达质粒,并通过脂质体转染并建立CCN3过表达的肝癌细胞株。利用transwell细胞侵袭实验和细胞划痕迁移实验检测CCN3对肝癌细胞侵袭迁移能力的影响。Western blot检测肝癌细胞株COX-2的表达变化来探讨相应机制。[结果]研究发现CCN3的RNA和蛋白水平在人类肝癌组织中均表达上调,并且其表达水平与临床肝癌患者的转移有一定相关性。各个肝癌细胞株间的CCN3表达有差异,但均高于人类正常肝细胞。成功建立了CCN3过表达的肝癌细胞株,并通过Western bolt验证CCN3蛋白表达水平。通过体外细胞功能实验transwell细胞侵袭实验和细胞划痕迁移实验证实了CCN3可以上调COX-2从而促进肝癌细胞的的侵袭与转移。[结论]CCN3在肝癌组织中表达异常,并可以通过COX-2表达的上调从而促进肝癌的的侵袭与转移,这些研究为CCN3将来成为肝癌预后指标及肝癌的分子靶向治疗提供了新的思路。
[Abstract]:[objective] CCN3 Nov Nephroblastoma overexpressed gene is a member of the CCN family and is involved in the development and development of many cell functions, including cell growth, differentiation, angiogenesis, adhesion and so on.At present, researchers have found that CCN3 is highly expressed in many kinds of malignant tumors and regulates the occurrence and development of many kinds of tumors, such as osteosarcoma, esophageal cancer and so on.However, the study of CCN3 in hepatocellular carcinoma has not been reported.In this study, we detected the expression of CCN3 in liver cancer tissues and cells, and carried out corresponding functional experiments to explore the corresponding mechanism.[methods] Real-time quantitative polymerase chain reaction (RT-PCR) assay and Western blot were used to detect the expression of CCN3 in hepatocellular carcinoma and its adjacent tissues.The correlation between the expression of CCN3 and the clinical data of patients with liver cancer was analyzed statistically.At the same time, the expression level of m RNA of CCN3 in the hepatoma cell line MHCC-97H SMMC-7721 Hep G2H2HH7MHCC-97L and normal human hepatocytes L02 was detected. The CCN3 overexpression plasmid was constructed, and the CCN3 overexpressed hepatoma cell line was transfected and established by liposome.Transwell cell invasion assay and cell scratch migration assay were used to detect the effect of CCN3 on the invasion and migration of hepatoma cells. Western blot was used to detect the expression of COX-2 in hepatoma cells.[results] it was found that the expression of RNA and protein of CCN3 was up-regulated in human liver cancer tissues, and its expression level was correlated with the metastasis of clinical HCC patients.The expression of CCN3 was higher than that of normal hepatocytes.CCN3 overexpression hepatoma cell line was successfully established, and the expression of CCN3 protein was verified by Western bolt.In vitro, transwell cell invasion assay and cell scratch migration assay confirmed that CCN3 can up-regulate COX-2 and promote the invasion and metastasis of hepatoma cells.[conclusion] the abnormal expression of CCN3 in HCC tissues can promote the invasion and metastasis of HCC through the up-regulation of COX-2 expression. These studies provide a new idea for CCN3 to be a prognostic indicator of HCC and a molecular targeted therapy for HCC in the future.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.7

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