ZNF750基因S70X突变体在食管鳞癌发生、发展中的功能研究

发布时间：2018-04-01 05:08

本文选题：食管癌　切入点：ZNF750　出处：《山西医科大学》2015年硕士论文

【摘要】：目的:研究锌指蛋白750(zinc finger protein 750,ZNF750)基因S70X突变体对食管鳞癌细胞生物学行为的影响。方法:在稳定敲除内源性ZNF750基因的KYSE150及KYSE140细胞中,转染野生型及突变型p LV-EGFP(2A)Puro-ZNF750 c DNA病毒载体,采用免疫荧光观察野生型及突变型ZNF750细胞内定位的差异,通过MTT实验、平板克隆形成实验、划痕实验和Transwell侵袭实验观察转染ZNF750突变体与转染野生型ZNF750相比,食管鳞癌细胞生物学行为的变化,PCR-Array检测转染ZNF750突变体与转染野生型ZNF750后基因表达量变化。结果:1、野生型ZNF750定位于细胞核,而S70X突变型ZNF750定位于胞浆。两者细胞内定位明显不同;2、ZNF750突变体S70X改变了野生型ZNF750引起的细胞增殖抑制作用,对食管癌细胞增殖、克隆形成、侵袭及迁移起促进作用;3、ZNF750突变体S70X与野生型ZNF750相比,肿瘤血管生成、上皮间质转化等相关基因表达水平存在显著差异。结论:S70X突变体改变了ZNF750基因生物学功能,发挥与野生型ZNF750相反的生物学作用,可能通过促进血管生成、上皮间质转化等促进食管癌细胞的发生发展。
[Abstract]:Objective: to study the effect of zinc finger protein 750(zinc finger protein 750G ZNF750) gene S70X mutant on the biological behavior of esophageal squamous cell carcinoma cells. Methods: in KYSE150 and KYSE140 cells with stable knockout of endogenous ZNF750 gene, the wild-type and mutant p#en5# c DNA virus vectors were transfected. The difference of intracellular localization between wild and mutant ZNF750 cells was observed by immunofluorescence. The transfection of ZNF750 mutants was compared with that of wild type ZNF750 by MTT test, plate clone formation test, scratch test and Transwell invasion experiment. Changes in biological behavior of esophageal squamous cell carcinoma cell lines: PCR-Array was used to detect the changes of gene expression after transfection of ZNF750 mutants and wild-type ZNF750. But S70X mutant ZNF750 was located in cytoplasm. The intracellular localization of S70X mutant S70X changed the inhibitory effect of wild-type ZNF750 on cell proliferation, proliferation and clone formation of esophageal carcinoma cells. Compared with wild type ZNF750, tumor angiogenesis, epithelial stromal transformation and other related gene expression levels were significantly different between S70X mutant S70X and wild type ZNF750. Conclusion the biological function of ZNF750 gene was changed by the mutant S70X. In contrast to wild-type ZNF750, it may promote the genesis and development of esophageal cancer cells by promoting angiogenesis and epithelium mesenchymal transformation.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1

【参考文献】