LIM家族蛋白Ajuba对肿瘤细胞的生物学功能与分子机制研究

发布时间：2018-01-15 03:04

本文关键词：LIM家族蛋白Ajuba对肿瘤细胞的生物学功能与分子机制研究　出处：《上海交通大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的探索结直肠癌细胞中Ajuba与Twist之间的相互作用,及该作用对于结直肠癌迁移所产生的影响。方法运用免疫共沉淀方法证明外源性Ajuba与Twist蛋白质之间存在相互作用;荧光素酶活性检测Ajuba对于Twist下游靶基因N-cadherin启动子的转录活性;构建结直肠癌SW1116-sh Ajuba/OEAjuba稳转细胞,并进行Transwell侵袭实验观察细胞迁移能力变化。结果Ajuba与Twsit蛋白质之间存在相互作用,并且Ajuba能通过Twist促进N-cadherin的表达调控。在结直肠癌细胞SW1116-sh Ajuba稳转细胞中可发现,AJUBA基因表达水平降低,蛋白表达下降,能抑制细胞迁移能力。结论Ajuba是转录因子Twist的共活化因子(coactivator),能够促进N-cadherin表达,增强结直肠癌细胞的迁移能力。目的探索Ajuba在雌激素受体α(Estrogen receptorα,ERα)阳性的乳腺癌细胞中的生物学作用,依据已知的Ajuba与ERα存在相互作用寻找涉及的分子机制。方法运用免疫共沉淀方法证明外源性Ajuba与ERα蛋白质之间存在相互作用;构建乳腺癌T47D-sh Ajuba、T47D-oe Ajuba稳转细胞,并进行CCK8实验观察细胞生长能力变化;建立T47D-sh ERα稳转细胞,比较生长趋势改变;荧光素酶活性检测Ajuba-ERα对于下游靶基因CDK6启动子的转录活性调控。结果Ajuba与ERα之间存在相互作用,并且Ajuba能通过ERα促进CDK6的表达调控。在乳腺癌细胞T47D-sh Ajuba稳转细胞中可发现,AJUBA基因表达水平降低,蛋白表达下降,细胞生长缓慢;细胞中过表达Ajuba,则可上调CDK6的表达水平。结论Ajuba是ERα的共活化因子(coactivator),能够促进CDK6表达,参与调控乳腺癌细胞T47D的生长。
[Abstract]:Objective to explore the interaction between Ajuba and Twist in colorectal cancer cells. Methods Immunoprecipitation method was used to prove the interaction between exogenous Ajuba and Twist protein. Luciferase activity was used to detect the transcriptional activity of Ajuba on N-cadherin promoter of Twist downstream gene. Construction of SW1116-sh Ajuba/OEAjuba stable transformed cells from colorectal cancer. The ability of cell migration was observed by Transwell invasion assay. Results there was interaction between Ajuba and Twsit protein. Ajuba can promote the regulation of N-cadherin expression through Twist. It can be found in SW1116-sh Ajuba stable transformed cells. The decrease of AJUBA gene expression and the decrease of protein expression can inhibit the ability of cell migration. Conclusion Ajuba is the coactivator of transcription factor Twist. It can promote N-cadherin expression and enhance the migration ability of colorectal cancer cells. Objective to explore the role of Ajuba in estrogen receptor 伪 strogen receptor 伪. The biological role of ER 伪 in breast cancer cells. Methods the interaction between exogenous Ajuba and ER 伪 protein was demonstrated by immunoprecipitation method. Breast cancer T47D-sh Ajubahe T47D-OE Ajuba was constructed, and the growth ability was observed by CCK8 assay. T47D-sh ER 伪 stable cells were established to compare the growth trend. Luciferase activity was used to detect the transcriptional regulation of Ajuba-ER 伪 on the downstream target gene CDK6 promoter. Results there was an interaction between Ajuba and ER 伪. Moreover, Ajuba can promote the regulation of CDK6 expression through ER 伪. The decrease of AJUBA gene expression can be found in T47D-sh Ajuba stable transfer cells of breast cancer cells. Protein expression decreased and cell growth slowed down. Overexpression of Ajuba can up-regulate the expression of CDK6. Conclusion Ajuba is a coactivator of ER 伪 and can promote the expression of CDK6. Participate in regulating the growth of breast cancer cell T47D.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.34

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