發(fā)布時(shí)間：2018-04-01 05:08

  本文選題：食管癌　切入點(diǎn)：ZNF750　出處：《山西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:研究鋅指蛋白750(zinc finger protein 750,ZNF750)基因S70X突變體對食管鱗癌細(xì)胞生物學(xué)行為的影響。方法:在穩(wěn)定敲除內(nèi)源性ZNF750基因的KYSE150及KYSE140細(xì)胞中,轉(zhuǎn)染野生型及突變型p LV-EGFP(2A)Puro-ZNF750 c DNA病毒載體,采用免疫熒光觀察野生型及突變型ZNF750細(xì)胞內(nèi)定位的差異,通過MTT實(shí)驗(yàn)、平板克隆形成實(shí)驗(yàn)、劃痕實(shí)驗(yàn)和Transwell侵襲實(shí)驗(yàn)觀察轉(zhuǎn)染ZNF750突變體與轉(zhuǎn)染野生型ZNF750相比,食管鱗癌細(xì)胞生物學(xué)行為的變化,PCR-Array檢測轉(zhuǎn)染ZNF750突變體與轉(zhuǎn)染野生型ZNF750后基因表達(dá)量變化。結(jié)果:1、野生型ZNF750定位于細(xì)胞核,而S70X突變型ZNF750定位于胞漿。兩者細(xì)胞內(nèi)定位明顯不同;2、ZNF750突變體S70X改變了野生型ZNF750引起的細(xì)胞增殖抑制作用,對食管癌細(xì)胞增殖、克隆形成、侵襲及遷移起促進(jìn)作用;3、ZNF750突變體S70X與野生型ZNF750相比,腫瘤血管生成、上皮間質(zhì)轉(zhuǎn)化等相關(guān)基因表達(dá)水平存在顯著差異。結(jié)論:S70X突變體改變了ZNF750基因生物學(xué)功能,發(fā)揮與野生型ZNF750相反的生物學(xué)作用,可能通過促進(jìn)血管生成、上皮間質(zhì)轉(zhuǎn)化等促進(jìn)食管癌細(xì)胞的發(fā)生發(fā)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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R735.1

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本文編號：1694097