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  本文選題：Pokemon + 胃癌��； 參考：《南華大學》2016年碩士論文

【摘要】：目的:觀察在Pokemon人胃癌細胞中對TGF-β1表達的影響及機制初步研究,分析其與臨床病理特征的相關性。方法:實驗分為兩部分。第一部分:1)以體外培養(yǎng)的MGC-803胃癌細胞系為研究對象,將Pokemon高表達質粒載體以及空載體質粒瞬時轉染MGC-803胃癌細胞系,先通過Q-PCR以及Western Blot檢測構建的高表達pokemon質粒,再通過熒光顯微鏡觀察過表達Pokemon對MGC-803胃癌細胞系中TGF-β1表達的影響,以及用Q-PCR檢測其表達情況,并通過Western Blot檢測Pokemon對部分EMT相關蛋白的表達影響;2)以體外培養(yǎng)的SGC-7901胃癌細胞系為研究對象,將Pokemon高表達質粒載體以及空載體質粒瞬時轉染SGC-7901胃癌細胞系,先通過Q-PCR以及Western Blot檢測構建的高表達pokemon質粒,再通過熒光顯微鏡觀察過表達Pokemon對SGC-7901胃癌細胞系中TGF-β1表達的影響,以及用Q-PCR檢測其表達情況,并通過Western Blot檢測Pokemon對部分EMT相關蛋白的表達影響;第二部分:應用雙熒光素酶報告基因活性實驗測定在MGC-803以及SGC-7901胃癌細胞系中轉染表達Pokemon質粒對TGF-β1啟動子的影響。結果:1)在MGC-803以及SGC-7901胃癌細胞系轉染Pokemon質粒后,通過Q-PCR以及Western Blot檢測到pokemon的表達水平增高,2)在MGC-803以及SGC-7901胃癌細胞系中,通過Q-PCR檢測到高表達Pokemon抑制TGF-β1的m RNA水平;3)在MGC-803以及SGC-7901胃癌細胞中,通過免疫熒光技術檢測到高表達Pokemon抑制TGF-β1表達;4)在MGC-803以及SGC-7901胃癌細胞系中,通過Western Blot檢測到高表達pokemon抑制人胃癌細胞中的部分EMT相關分子的表達;5)在MGC-803以及SGC-7901胃癌細胞系中,通過雙熒光素酶報告基因活性實驗檢測到高表達Pokemon通過抑制TGF-β1的啟動子從而影響其表達。結論:1)Pokemon抑制MGC-803以及SGC-7901胃癌細胞系中EMT的發(fā)生,其通過抑制TGF-β1的表達從而調節(jié)EMT過程;2)Pokemon在胃癌細胞中抑制TGF-β1的表達,其機制和Pokemon通過抑制TGF-β1的啟動子活性有關。
[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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R735.2

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本文編號：1950845