REGγ促进人乳腺癌MDA-MB-231细胞株发生上皮—间充质转化

发布时间：2018-03-21 05:27

本文选题：REGγ　切入点：MDA-MB-231　出处：《重庆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:研究REGγ基因促进人乳腺癌细胞株MDA-MB-231发生上皮-间充质转化的生物学作用。方法:免疫组化检测乳腺癌组织与癌旁组织REGγ的表达差异,以及检测转移淋巴结中REGγ的表达水平。Western Blot和RT-PCR检测不同人乳腺癌细胞系之间REGγ的表达差异。构建REGγ基因过表达和干扰载体并将其导入MDA-MB-231细胞中,然后用RT-PCR和细胞免疫荧光分别从转录水平和蛋白水平上检测上皮-间充质转化细胞标记物的表达水平的变化。通过划痕实验和Transwell阐释REGγ对细胞侵袭迁移能力的影响。结果:免疫组化结果提示REGγ在乳腺癌组织和转移淋巴结中显著高表达。Western Blot和RT-PCR提示REGγ在MDA-MB-231细胞中表达水平较高。MDA-MB-231细胞过表达REGγ后E-钙粘素(E-cadherin)下调,而波形蛋白(Vimentin)上调(P0.01);干扰REGγ后会逆转上皮-间充质转化,使细胞侵袭迁移能力减弱。结论:REGγ促进人乳腺癌MDA-MB-231细胞发生上皮-间充质转化,从而促使肿瘤细胞发生侵袭转移。
[Abstract]:Objective: to study the biological effect of REG 纬 gene on epithelial-mesenchymal transformation in human breast cancer cell line MDA-MB-231. The expression level of REG 纬 in metastatic lymph nodes was detected. Western Blot and RT-PCR were used to detect the difference of REG 纬 expression between different human breast cancer cell lines. The overexpression and interference vector of REG 纬 gene was constructed and introduced into MDA-MB-231 cells. Then the expression level of epithelium-mesenchymal transformation cell marker was detected by RT-PCR and cellular immunofluorescence at the transcriptional level and protein level respectively. The effect of REG 纬 on the ability of cell invasion and migration was explained by scratch test and Transwell. Results: immunohistochemical results showed that REG 纬 was significantly overexpressed in breast cancer tissues and metastatic lymph nodes. Western Blot and RT-PCR suggested that REG 纬 expression was higher in MDA-MB-231 cells. Ecadherin E-cadherin was down-regulated in MDA-MB-231 cells after overexpression of REG 纬. Vimentin upregulated P0.01C, and interfered with REG 纬, which reversed the epithelial-mesenchymal transformation and weakened the ability of invasion and migration. Conclusion: Vimentin 纬 can promote the epithelial-mesenchymal transformation of human breast cancer MDA-MB-231 cells and promote the invasion and metastasis of tumor cells.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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