PARD6A在卵巢癌中的作用机制研究

发布时间：2018-01-18 07:06

本文关键词：PARD6A在卵巢癌中的作用机制研究　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：分割缺陷蛋白6同源物α(Partitioning defective 6 homolog alpha,PARD6A)基因目前的研究主要聚焦在它作为细胞极化和细胞不对称分裂的一个重要的分子支架蛋白的作用,尽管日前已有少量文献报道了该基因可能与乳腺癌及肺癌的发生发展有关[1,2]。并且多篇文献报道,极性蛋白很有可能影响肿瘤的迁移和侵润。这个基因在卵巢癌中还未曾研究过,对这个基因在卵巢癌的作用和机制的研究,有可能定义出全新的卵巢癌靶向基因,用以启动新的靶向药物研究。因此,以PARD6A基因为目标,实验首先考察PARD6A基因与卵巢癌的相关性。通过特异性沉默或过表达PARD6A基因,检测了该基因对卵巢癌细胞株功能的影响,此外,进一步研究了PARD6A基因的蛋白表达量与临床卵巢癌肿瘤相关性,在探索了临床相关的基础上,进一步研究了PARD6A在卵巢癌中的作用机制。经免疫印迹技术检测到,PARD6A基因在浆液性卵巢癌细胞株SKOV3和A2780是异常高表达的;经免疫组织化学技术检测到PARD6A基因的蛋白在浆液性和粘液性卵巢肿瘤临床组织样本中高表达(其中浆液性是卵巢癌中最常见的组织学类型),比在正常卵巢组织中的表达显著高。经细胞划痕实验、Transwell小室法、吸附实验及免疫荧光技术等检测到,沉默PARD6A基因,抑制了卵巢癌细胞株迁移、侵润能力和减弱吸附能力等;同时,PARD6A过量表达促进了锚定非依赖性生长、细胞的迁移和侵润等。经卵巢癌的小鼠肺转移模型实验,验证出以小发卡RNA(short hairpin RNA,shRNA)沉默PARD6A基因,可以在小鼠体内能够抑制卵巢癌细胞SKOV3的转移;对于PARD6A对卵巢癌抑制迁移和侵润的机制,本课题初步检测到PARD6A基因可能影响了Cdc42和RhoA介导的细胞迁移和侵润相关的信号通路。综上所述,本课题旨在考察PARD6A基因在卵巢癌的作用和机制,希望因此能发现全新的卵巢癌靶向基因,用以改善卵巢癌的治疗和预后,防止肿瘤的恶化和复发。
[Abstract]:The fragment defect protein 6 congener 伪 -partitioning defective 6 homolog alpha. The current study of the PARD6A) gene focuses on its role as an important molecular scaffold protein for cell polarization and asymmetric cell division. Although a small amount of literature has reported that the gene may be related to the occurrence and development of breast cancer and lung cancer. [The polarity protein may affect tumor migration and invasion. This gene has not been studied in ovarian cancer, and the role and mechanism of this gene in ovarian cancer have been studied. It may be possible to define new ovarian cancer targeting genes to initiate new targeted drug research. Therefore, target the PARD6A gene. Firstly, the relationship between PARD6A gene and ovarian cancer was investigated. The effect of PARD6A gene on ovarian cancer cell line function was detected by specific silencing or overexpression of PARD6A gene. The relationship between the protein expression of PARD6A gene and clinical ovarian cancer was further studied. The mechanism of PARD6A in ovarian cancer was further studied and detected by Western blotting. The expression of PARD6A gene was abnormally high in serous ovarian cancer cell lines SKOV3 and A2780. The protein of PARD6A gene was detected by immunohistochemistry in the clinical samples of serous and mucinous ovarian tumors (serous is the most common histological type of ovarian cancer). The expression of PARD6A gene was significantly higher than that in normal ovarian tissue. The PARD6A gene was silenced by cell scratch assay, transwell chamber assay, adsorption assay and immunofluorescence technique. Inhibition of ovarian cancer cell line migration, invasion ability and reduced adsorption ability, etc. At the same time, the overexpression of PARD6A promoted anchoring independent growth, cell migration and invasion. The silencing of PARD6A gene with small hairpin RNA(short hairpin RNAs was verified. The SKOV3 metastasis of ovarian cancer cells could be inhibited in mice. The mechanism of inhibition of migration and invasion of ovarian cancer by PARD6A. In this study, we preliminarily detected that PARD6A gene may affect Cdc42 and RhoA mediated cell migration and invasion signal pathway. The purpose of this study is to investigate the role and mechanism of PARD6A gene in ovarian cancer, so as to find a novel target gene for ovarian cancer in order to improve the treatment and prognosis of ovarian cancer and prevent the deterioration and recurrence of ovarian cancer.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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