Wipl调控浆液性卵巢癌细胞增殖以及转移侵袭功能及其与临床相关性初步研究

发布时间：2018-04-02 10:53

本文选题：wip1　切入点：浆液性卵巢癌　出处：《复旦大学》2014年硕士论文

【摘要】：目的研究野生型p53诱导的蛋白磷酸酶1(wip1)对卵巢癌细胞增殖以及转移侵袭的调控功能,并检测wip1在浆液性卵巢癌组织中的表达水平,分析其与生存的相关性。方法1.构建wipl沉默和过表达卵巢癌细胞系,检测其细胞内wip1蛋白及mRNA水平变化;分析其细胞周期、克隆形成能力以及细胞增殖情况的变化；通过划痕实验和Transwell实验检测其对细胞转移和侵袭能力的影响；使用免疫荧光及Western blot方法检测wip1及其相关蛋白的变化情况。2.收集本院初次手术的高级别浆液性卵巢癌组织150例以及正常卵巢组织10例,使用免疫组织化学方法检测wip1在高级别浆液性卵巢癌中的表达情况,同时搜集相关病理、手术及随访信息,统计分析其与预后的关系。结果1.在wip1 shRNA卵巢癌细胞中,wip1的mRNA及蛋白水平明显低于对照组,而在wip1 cDNA卵巢癌细胞中,Wip1的mRNA及蛋白水平明显高于对照组。wip1表达被干扰后,细胞阻滞于G1期；细胞生长速度减慢、克隆数量减少。在转移相关实验发现,Wip1沉默后细胞的迁移和侵袭性增强；转移相关蛋白E-cadherin水平降低,而N-cadherin、vimentin、MMP9、VEGF等蛋白的表达水平有不同程度的升高,转录因子slug和snail表达水平也升高。2.免疫组化结果显示wip1在高级别浆液性卵巢癌组织中呈现不同程度的阳性表达；临床因素相关性分析,wip1的表达程度在肿瘤的药物敏感性存在明显差异(p=0.036),而与年龄、病理分期、腹水量无相关性。wipl强阳性表达患者的无进展生存期和总生存期均明显低于弱阳性表达者,其差异具有统计学意义。结论wip1在浆液性卵巢癌细胞增殖和转移过程中发挥作用,并且可能与EMT相关。在高级别浆液性卵巢癌组织中wip1存在不同程度的阳性表达,wipl的表达程度与患者对铂类药物的敏感性及预后相关。
[Abstract]:Objective to investigate the effects of wild-type p53 induced protein phosphatase 1 (p3p1) on the proliferation, metastasis and invasion of ovarian cancer cells, and to detect the expression of wip1 in serous ovarian carcinoma, and to analyze its correlation with survival.Method 1.Wipl silencing and overexpression of ovarian cancer cell lines were constructed to detect the changes of wip1 protein and mRNA levels, cell cycle, clone formation ability and cell proliferation were analyzed.The effects on cell metastasis and invasion were detected by scratch test and Transwell assay, and the changes of wip1 and its associated proteins were detected by immunofluorescence and Western blot.150 cases of high grade serous ovarian carcinoma and 10 cases of normal ovarian tissue were collected from our hospital for the first time. Immunohistochemical method was used to detect the expression of wip1 in high grade serous ovarian cancer.The relationship between surgical and follow-up information and prognosis was analyzed statistically.Result 1.The levels of mRNA and protein in wip1 shRNA ovarian cancer cells were significantly lower than those in the control group, while the mRNA and protein levels of Wip1 in wip1 cDNA ovarian cancer cells were significantly higher than those in the control group.The number of clones decreased.In the metastasis related experiment, we found that the cell migration and invasiveness were enhanced after silencing of Wip1, the E-cadherin level of metastasis related protein decreased, but the expression level of N-cadherinvimentinMMP9 protein increased in varying degrees, and the expression level of transcription factor slug and snail was also increased. 2.Immunohistochemical results showed that the expression of wip1 in high grade serous ovarian carcinoma was positive in different degrees, the correlation analysis of clinical factors showed that there were significant differences in the drug sensitivity of the tumor, but with age and pathological stage.The progression-free survival time and total survival time of the patients with strong positive expression of ascites were significantly lower than those of the patients with weak positive expression, and the difference was statistically significant.Conclusion wip1 may play a role in the proliferation and metastasis of ovarian cancer cells and may be related to EMT.The positive expression of wip1 in high grade serous ovarian cancer tissues was correlated with the sensitivity and prognosis of patients with platinum.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31

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