本文選題：ABCG2　切入點(diǎn)：基因表達(dá)　出處：《石河子大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過檢測過表達(dá)ABCG2的細(xì)胞模型砷耐受性的改變,初步探討ABCG2與抗砷性產(chǎn)生關(guān)系。 方法:(1)免疫熒光化學(xué)方法檢測ChAsE-BMSCs、膀胱癌抗砷細(xì)胞以及23例急性白血病患者骨髓內(nèi)ABCG2的表達(dá)情況;(2)真核表達(dá)載體pEGFP-C1-ABCG2經(jīng)轉(zhuǎn)化、篩選、測序鑒定后,抽提并獲取高純度的重組質(zhì)粒,采用脂質(zhì)體介導(dǎo)轉(zhuǎn)染法轉(zhuǎn)染至293T細(xì)胞,western-blot鑒定轉(zhuǎn)染效果(3)臺盼蘭計(jì)數(shù)方法檢測ABCG2過表達(dá)對293T細(xì)胞增殖的影響(4)細(xì)胞毒性實(shí)驗(yàn)檢測ABCG2過表達(dá)細(xì)胞對砷劑耐受性的改變。 結(jié)果: (1)免疫熒光方法檢測兩種抗砷細(xì)胞中ABCG2蛋白表達(dá)均低于對照組細(xì)胞;(2)免疫熒光方法檢測23例急性白血病患者ABCG2蛋白表達(dá)強(qiáng)度,從低到高依次為:正常對照組、初發(fā)組和復(fù)發(fā)組,三者之間差異有統(tǒng)計(jì)學(xué)意義(P0.0001)(3)激光共聚焦顯微鏡觀察和Western-blot結(jié)果證明轉(zhuǎn)染成功;(4)ABCG2過表達(dá)降低細(xì)胞的增殖能力;(5)ABCG2過表達(dá)細(xì)胞對砷劑耐受性下降。 結(jié)論: (1)抗砷細(xì)胞內(nèi)ABCG2蛋白表達(dá)下調(diào),細(xì)胞抗砷性與ABCG2表達(dá)呈負(fù)相關(guān);(2)ABCG2高表達(dá)是急性白血病患者不利的預(yù)后因素;(3)成功建立過表達(dá)ABCG2基因的細(xì)胞模型;
[Abstract]:Aim: to investigate the relationship between ABCG2 and arsenic resistance by detecting the changes of arsenic tolerance in a cell model with overexpression of ABCG2. Methods the expression of ChAsE-BMSCs, anti-arsenic cells of bladder cancer and bone marrow ABCG2 of 23 patients with acute leukemia were detected by the immunofluorescence method. The eukaryotic expression vector pEGFP-C1-ABCG2 was transformed, screened and sequenced, and the recombinant plasmid was extracted and obtained. The effects of ABCG2 overexpression on the proliferation of 293T cells were evaluated by liposome-mediated transfection into 293T cells by western-blot. The effects of ABCG2 overexpression on the proliferation of 293T cells were detected by Trypan blue counting method. The changes of arsenic tolerance of ABCG2 overexpression cells were detected by cytotoxicity assay. Results: the expression of ABCG2 protein in two kinds of arsenic resistant cells was lower than that in control group (P < 0.01). Immunofluorescence method was used to detect the expression of ABCG2 protein in 23 patients with acute leukemia. The order from low to high was normal control group. The results of laser confocal microscopy and Western-blot showed that the overexpression of ABCG2 decreased the proliferative ability of the cells. Conclusion: (1) the expression of ABCG2 in arsenic resistant cells is down-regulated, and the high expression of ABCG2 is negatively correlated with the expression of ABCG2. The high expression of ABCG2 is a negative prognostic factor in patients with acute leukemia. A cell model of overexpression of ABCG2 gene is successfully established.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R346

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

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本文編號：1575765