MiR-126调控cofilin-1与上皮性卵巢癌侵袭转移关系的研究

发布时间：2017-12-31 01:22

本文关键词：MiR-126调控cofilin-1与上皮性卵巢癌侵袭转移关系的研究　出处：《浙江大学》2016年硕士论文　论文类型：学位论文

更多相关文章： 卵巢癌 侵袭 miR-126 Cofilin-1

【摘要】：研究背景和目的：卵巢癌因其进展迅速,容易腹腔内广泛转移,因此在确诊时多数患者已处于晚期,目前已经成为妇科致死率最高的肿瘤之一。卵巢癌细胞恶性生物学行为强,大部分患者接受标准化治疗后2年左右出现肿瘤复发及耐药,因此晚期卵巢癌患者5年生存率较低。近年来研究表明各种miRNA可通过作用于]mRNA转录后水平在肿瘤发生发展中起到抑癌或促癌作用。对于miR-126,有研究表明在乳腺癌、肺癌、前列腺癌等多种肿瘤发生发展中起着抑癌基因的作用,但其在卵巢癌中的作用尚不明确。丝切蛋白(Cofilin)是一种肌动蛋白结合蛋白,在细胞骨架重塑和移动调节上起着重要作用,而卵巢癌等恶性肿瘤细胞的分裂、迁移及特异性结构依赖于细胞骨架的动力学调节。为此本研究拟通过体外培养SKOV3卵巢癌细胞株并转染不同miR-126基因片段,比较miR-126与Cofilia-1表达之间的关系,以此研究miR-126是否可通过调控conflin-1而参与卵巢癌细胞的侵袭转移的调节。研究材料和方法：本研究将人卵巢浆液性囊腺癌SKOV3细胞株进行体外培养,miR-126通过慢病毒的包装后转染人卵巢浆液性囊腺癌SKOV3细胞系,上述卵巢癌细胞分成四组,分别为：miR-126过表达组(SKOV3+LV3-has-miR-126组)、miR-126抑制表达组(SKOV3+LV3-has-miR-126 inhibitor组)、转染阴性对照组(SKOV3+LV3NC组)和空白对照组(SKOV3组),实验采用Transwell侵袭实验观察各组细胞的侵袭能力,通过免疫荧光法观察cofilin-1表达水平的差异。结果：1.Transwell侵袭实验的结论表明miR-126过表达组(SKOV3+LV3-has-miR-126组SKOV3细胞侵犯基质胶的穿膜细胞数明显少于miR-126抑制表达组(SKOV3+LV3-has-miR-126 inhibitor组)(P0.05),同时也明显少于转染阴性对照组(SKOV3+LV3NC组)和空白对照组(SKOV3组)的穿膜细胞数(P均0.05)。2.免疫荧光检测结果显示,LV3-has-miR-126组细胞Cofilin-1表达明显低于miR-126抑制表达组(SKOV3+LV3-has-miR-126 inhibitor组)、转染阴性对照组(SKOV3+LV3NC组)和空白对照组(SKOV3组),而LV3-has-miR-126 inhibitor组Cofilin-1表达明显高于转染阴性对照组(SKOV3+LV3NC组)和空白对照组(SKOV3组)。结论：1. miR-126过表达可以减弱SKOV3细胞侵袭能力,提示miR-126过表达可能抑制卵巢癌细胞的侵袭转移。2.转染miR-126的SKOV3细胞cofilin-1表达明显减少,提示miR-126可能通过抑制cofil in-1表达,进而影响细胞骨架而抑制卵巢癌细胞的侵袭转移。
[Abstract]:Background and objective: ovarian cancer is in advanced stage because of its rapid progression and easy metastasis in abdominal cavity. Ovarian cancer cells have strong malignant biological behavior, most of the patients received standardized treatment about 2 years after the tumor recurrence and drug resistance. Therefore, the 5-year survival rate of patients with advanced ovarian cancer is low. In recent years, studies have shown that various kinds of miRNA can inhibit or promote cancer in tumorigenesis and development by acting on] mRNA posttranscriptional level. -126. Studies have shown that breast cancer, lung cancer, prostate cancer and other tumors play a role in the development of tumor suppressor genes. However, its role in ovarian cancer is unclear. Cofilin is an actin binding protein, which plays an important role in cytoskeletal remodeling and movement regulation. And the division of malignant tumor cells such as ovarian cancer. Migration and specific structure depend on the dynamics of cytoskeleton. This study aims to culture SKOV3 ovarian cancer cell lines in vitro and transfect different miR-126 gene fragments. To compare the relationship between miR-126 and Cofilia-1 expression. To study whether miR-126 is involved in the regulation of invasion and metastasis of ovarian cancer cells by regulating conflin-1. Materials and methods:. In this study, human ovarian serous cystadenocarcinoma cell line SKOV3 was cultured in vitro. MiR-126 was transfected into human ovarian serous cystadenocarcinoma SKOV3 cell line after packaging of lentivirus. The ovarian cancer cells were divided into four groups. SKOV3 LV3-has-miR-126 group was the overexpressed group of 10% miR-126 (P < 0.05). MiR-126 inhibited expression of SKOV3 LV3-has-miR-126 inhibitor. SKOV3 LV3NC (negative control group) and SKOV3 (blank control group) were transfected. Transwell invasion assay was used to observe the invasiveness of the cells in each group. The expression of cofilin-1 was observed by immunofluorescence method. Results 1. The results of Transwell invasion experiment showed that the miR-126 overexpression group (. The number of SKOV3 cells invading matrix glue in SKOV3 LV3-has-miR-126 group was significantly less than that in miR-126 inhibited expression group (2). SKOV3 LV3-has-miR-126 inhibitor group (P0.05). At the same time, the number of perforating cells in SKOV3 LV3NC group (negative transfection group) and SKOV3 group (blank control group) were significantly lower than that in SKOV3 group. The expression of Cofilin-1 in LV3-has-miR-126 group was significantly lower than that in miR-126 group (. SKOV3 LV3-has-miR-126 inhibitor group. Transfection negative control group (SKOV3 LV3NC group) and blank control group (SKOV3 group). The expression of Cofilin-1 in LV3-has-miR-126 inhibitor group was significantly higher than that in negative control group (SKOV3 LV3NC group) and blank control group (P < 0.05). Conclusion: 1. Overexpression of miR-126 can attenuate the invasiveness of SKOV3 cells. These results suggest that the overexpression of miR-126 may inhibit the invasion and metastasis of ovarian cancer cells. 2. The expression of cofilin-1 in SKOV3 cells transfected with miR-126 was significantly decreased. The results suggest that miR-126 inhibits the invasion and metastasis of ovarian cancer cells by inhibiting the expression of cofil in-1 and thus affecting the cytoskeleton.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R737.31

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