Pokemon在胃癌细胞中对TGF-β1表达的影响及机制初步研究

发布时间：2018-05-29 12:15

本文选题：Pokemon + 胃癌　；参考：《南华大学》2016年硕士论文

【摘要】：目的:观察在Pokemon人胃癌细胞中对TGF-β1表达的影响及机制初步研究,分析其与临床病理特征的相关性。方法:实验分为两部分。第一部分:1)以体外培养的MGC-803胃癌细胞系为研究对象,将Pokemon高表达质粒载体以及空载体质粒瞬时转染MGC-803胃癌细胞系,先通过Q-PCR以及Western Blot检测构建的高表达pokemon质粒,再通过荧光显微镜观察过表达Pokemon对MGC-803胃癌细胞系中TGF-β1表达的影响,以及用Q-PCR检测其表达情况,并通过Western Blot检测Pokemon对部分EMT相关蛋白的表达影响;2)以体外培养的SGC-7901胃癌细胞系为研究对象,将Pokemon高表达质粒载体以及空载体质粒瞬时转染SGC-7901胃癌细胞系,先通过Q-PCR以及Western Blot检测构建的高表达pokemon质粒,再通过荧光显微镜观察过表达Pokemon对SGC-7901胃癌细胞系中TGF-β1表达的影响,以及用Q-PCR检测其表达情况,并通过Western Blot检测Pokemon对部分EMT相关蛋白的表达影响;第二部分:应用双荧光素酶报告基因活性实验测定在MGC-803以及SGC-7901胃癌细胞系中转染表达Pokemon质粒对TGF-β1启动子的影响。结果:1)在MGC-803以及SGC-7901胃癌细胞系转染Pokemon质粒后,通过Q-PCR以及Western Blot检测到pokemon的表达水平增高,2)在MGC-803以及SGC-7901胃癌细胞系中,通过Q-PCR检测到高表达Pokemon抑制TGF-β1的m RNA水平;3)在MGC-803以及SGC-7901胃癌细胞中,通过免疫荧光技术检测到高表达Pokemon抑制TGF-β1表达;4)在MGC-803以及SGC-7901胃癌细胞系中,通过Western Blot检测到高表达pokemon抑制人胃癌细胞中的部分EMT相关分子的表达;5)在MGC-803以及SGC-7901胃癌细胞系中,通过双荧光素酶报告基因活性实验检测到高表达Pokemon通过抑制TGF-β1的启动子从而影响其表达。结论:1)Pokemon抑制MGC-803以及SGC-7901胃癌细胞系中EMT的发生,其通过抑制TGF-β1的表达从而调节EMT过程;2)Pokemon在胃癌细胞中抑制TGF-β1的表达,其机制和Pokemon通过抑制TGF-β1的启动子活性有关。
[Abstract]:Aim: to investigate the effect of TGF- 尾 1 expression in Pokemon human gastric cancer cells and its mechanism, and to analyze the correlation between TGF- 尾 1 expression and clinicopathological features. Methods: the experiment was divided into two parts. In the first part, the MGC-803 gastric cancer cell line cultured in vitro was used as the research object. The Pokemon high expression plasmid vector and the empty body mass were transient transfected into the MGC-803 gastric cancer cell line. The high expression pokemon plasmid was constructed by Q-PCR and Western Blot detection. The effect of overexpression of Pokemon on the expression of TGF- 尾 1 in MGC-803 gastric cancer cell line was observed by fluorescence microscope, and the expression of TGF- 尾 1 was detected by Q-PCR. The effect of Pokemon on the expression of some EMT related proteins was detected by Western Blot. The high expression plasmid vector of Pokemon and the unloaded body mass were transiently transfected into SGC-7901 gastric cancer cell line using SGC-7901 gastric cancer cell line cultured in vitro. The high expression pokemon plasmid was constructed by Q-PCR and Western Blot detection, then the effect of overexpression Pokemon on the expression of TGF- 尾 1 in SGC-7901 gastric cancer cell line was observed by fluorescence microscope, and the expression of TGF- 尾 1 in SGC-7901 gastric cancer cell line was detected by Q-PCR. The effect of Pokemon on the expression of some EMT related proteins was detected by Western Blot. Part two: the effect of transfection and expression of Pokemon plasmid on TGF- 尾 1 promoter in MGC-803 and SGC-7901 gastric cancer cell lines was determined by double luciferase reporter gene activity assay. Results after MGC-803 and SGC-7901 gastric cancer cell lines were transfected with Pokemon plasmid, the expression of pokemon was detected by Q-PCR and Western Blot. High expression of Pokemon inhibited the expression of TGF- 尾 1 in MGC-803 and SGC-7901 gastric cancer cells, and high expression of Pokemon inhibited TGF- 尾 1 expression in MGC-803 and SGC-7901 gastric cancer cell lines by immunofluorescence technique. Overexpression of pokemon could inhibit the expression of some EMT related molecules in human gastric cancer cells by Western Blot) in MGC-803 and SGC-7901 gastric cancer cell lines. The high expression of Pokemon was detected by double luciferase reporter gene activity assay, and the expression of TGF- 尾 1 was inhibited by inhibiting the promoter of TGF- 尾 1. Conclusion the expression of TGF- 尾 1 in gastric cancer cells is inhibited by the inhibition of EMT in MGC-803 and SGC-7901 gastric cancer cell lines by the inhibition of the expression of TGF- 尾 1. The mechanism of the inhibition of TGF- 尾 1 expression by Pokemon is related to the inhibition of the promoter activity of TGF- 尾 1 by inhibiting the expression of TGF- 尾 1 in gastric cancer cells by inhibiting the expression of TGF- 尾 1.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.2

【相似文献】