GOLPH3在卵巢癌组织中的表达及作用机制的初步研究

发布时间：2018-03-12 09:26

本文选题：卵巢癌　切入点：GOLPH3　出处：《泰山医学院》2014年硕士论文　论文类型：学位论文

【摘要】：背景卵巢癌（ovarian carcinoma，OC）是女性生殖系统常见的恶性肿瘤，早期不易发现，一旦发现常常已属晚期。上皮性卵巢癌（Epithelial Ovarian Cancer, EOC)是卵巢癌中最常见的一种类型，是妇科肿瘤中死亡率最高的一种类型。目前卵巢癌的常规治疗仍然是手术、化疗、放疗和内分泌治疗等措施，但这些治疗方法都存在着疗效不佳的问题。随着分子生物学的发展，肿瘤分子的靶向治疗成为众多学者关注的热点，，基因干扰技术正逐步用于目的基因及相关基因的作用机制的研究。高尔基体磷蛋白3（GOLPH3）是近年来发现的一种癌基因，在多种实体肿瘤中存在过表达，并与肿瘤的临床分期、肿瘤病理分级及转移侵袭密切相关。GOLPH3在多种生物学过程中发挥作用，参与多种恶性肿瘤的发展过程，其高表达多与不良的临床预后有关，而GOLPH3 在卵巢癌的发生、发展中的作用机制目前还不清楚。目的本课题旨在研究GOLPH3在卵巢浆液性囊腺癌组织中的表达及其与临床病理特征之间的关系，并利用基因干扰技术初步探讨GOLPH3在卵巢癌细胞生物学行为方面的的作用机制，阐明GOLPH3在卵巢癌发生、发展过程中的作用。方法 1.应用实时荧光定量PCR、蛋白印记法(Western blot)分别检测各组织标本中GOLPH3在mRNA和蛋白水平的表达，并与患者临床病理特征进行相关性分析。 2.构建针对人GOLPH3基因的shRNA表达质粒（GOLPH3-shRNA），转染人卵巢癌SW626细胞系，使GOLPH3基因沉默。 3.利用CCK-8、细胞克隆形成实验及Transwell小室法检测GOLPH3-shRNA转染后对SW626细胞增殖、迁移及侵袭能力的影响。结果 1.GOLPH3mRNA在卵巢浆液性囊腺癌组织中的表达高于卵巢囊腺瘤组（2.33-7.34倍），差异具有统计学意义（P＜0.05）。GOLPH3蛋白在卵巢浆液性囊腺癌组织中阳性表达率（73.8%）明显高于卵巢囊腺瘤组（35.71%）（P＜0.01），差异具有统计学意义。 2.GOLPH3蛋白阳性表达与浆液性囊腺癌的临床分期(p=0.019)、病理分级(p=0.042)及有无淋巴结转移(p=0.000)有关，而与患者的年龄（p=0.881）无关（P0.05）；并且肿瘤细胞的分化程度越低其GOLPH3蛋白表达量越高（P＜0.01），差异具有统计学意义。 3.细胞实验中干扰质粒转染48小时后SW626细胞的转染效率约为75%，干扰组的GOLPH3的mRNA及蛋白表达水平的表达均明显下调(P 0.01)，差异具有统计学意义。 4.干扰组肿瘤细胞的增殖能力受抑制(P 0.01)，且这种抑制作用与时间无关；卵巢癌肿瘤细胞的克隆形成能力降低(P 0.01)；卵巢癌肿瘤细胞的转移及侵袭能力亦明显降低(P 0.01)，差异具有统计学意义。结论 1．GOLPH3在卵巢浆液性囊腺癌的发生、发展中发挥着重要的作用。 2．GOLPG3促进卵巢癌的增殖、生长、迁移及侵袭，有可能成为卵巢癌治疗新的靶基因。
[Abstract]:Background. Ovarian carcinoma carcinoma OCis a common malignant tumor in the female reproductive system. It is not easy to detect early, and is often late once found. Epithelial ovarian cancer Epithelial Ovarian carcinoma (EOC) is the most common type of ovarian cancer. It is one of the highest mortality rates among gynecological tumors. At present, conventional treatment for ovarian cancer is still surgery, chemotherapy, radiotherapy and endocrine therapy. With the development of molecular biology, the targeted therapy of tumor molecules has become the focus of attention of many scholars. Gene interference technique is gradually being used to study the action mechanism of target gene and related gene. Golgi body phosphoprotein 3 (Golgi) is a kind of oncogene found in recent years, which is overexpressed in many solid tumors and is associated with the clinical stage of tumor. GOLPH3 plays a role in many biological processes and participates in the development of many kinds of malignant tumors. The high expression of GOLPH3 is related to the poor clinical prognosis. The mechanisms underlying the development of ovarian cancer are unclear. Purpose. The purpose of this study was to investigate the expression of GOLPH3 in ovarian serous cystadenocarcinoma and its relationship with clinicopathological features, and to explore the role of GOLPH3 in the biological behavior of ovarian cancer cells by gene interference technique. To elucidate the role of GOLPH3 in the genesis and development of ovarian cancer. Method. 1. The expression of GOLPH3 in mRNA and protein was detected by real-time fluorescence quantitative PCR and Western blot, and the correlation was analyzed with the clinicopathological features of the patients. 2. The shRNA expression plasmid of human GOLPH3 gene was constructed and transfected into human ovarian cancer SW626 cell line. The GOLPH3 gene was silenced. 3. The effects of GOLPH3-shRNA transfection on the proliferation, migration and invasion of SW626 cells were detected by CCK-8, cell clone formation assay and Transwell chamber assay. Results. 1. The expression of GOLPH3 mRNA in ovarian serous cystadenocarcinoma was 2.33-7.34 times higher than that in ovarian cystadenoma (P < 0.05). The positive expression rate of GOLPH3 protein in ovarian serous cystadenocarcinoma was significantly higher than that in ovarian cystadenoma (35.71, P < 0.01, P < 0.01), and the positive expression rate of GOLPH3 protein in ovarian serous cystadenocarcinoma was significantly higher than that in ovarian cystadenoma (P < 0.05). It has statistical significance. 2. The positive expression of GOLPH3 protein was related to the clinical stage of serous cystadenocarcinoma, p0. 019, pathological grade p0. 042) and lymph node metastasis (p0. 000). And the lower the differentiation degree of tumor cells, the higher the expression of GOLPH3 protein (P < 0.01). 3. In the cell experiment, the transfection efficiency of SW626 cells was about 75 after 48 hours of interference plasmid transfection. The expression of mRNA and protein of GOLPH3 in the interference group was significantly down-regulated (P 0.01), and the difference was statistically significant. 4. The proliferation ability of tumor cells in the interference group was inhibited by P0.01, and the inhibitory effect was not related to time. The clone forming ability of ovarian cancer cells decreased P 0.01, and the metastasis and invasion ability of ovarian cancer cells decreased significantly (P 0.01), the difference was statistically significant. Conclusion. GOLPH3 plays an important role in the development of ovarian serous cystadenocarcinoma. 2. GOLPG3 promotes the proliferation, growth, migration and invasion of ovarian cancer and may become a new target gene for ovarian cancer therapy.
【学位授予单位】：泰山医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31

【相似文献】