ABCG2与抗砷性产生关系的研究

发布时间：2018-03-06 17:21

本文选题：ABCG2　切入点：基因表达　出处：《石河子大学》2009年硕士论文　论文类型：学位论文

【摘要】：目的:通过检测过表达ABCG2的细胞模型砷耐受性的改变,初步探讨ABCG2与抗砷性产生关系。方法:(1)免疫荧光化学方法检测ChAsE-BMSCs、膀胱癌抗砷细胞以及23例急性白血病患者骨髓内ABCG2的表达情况;(2)真核表达载体pEGFP-C1-ABCG2经转化、筛选、测序鉴定后,抽提并获取高纯度的重组质粒,采用脂质体介导转染法转染至293T细胞,western-blot鉴定转染效果(3)台盼兰计数方法检测ABCG2过表达对293T细胞增殖的影响(4)细胞毒性实验检测ABCG2过表达细胞对砷剂耐受性的改变。结果: (1)免疫荧光方法检测两种抗砷细胞中ABCG2蛋白表达均低于对照组细胞;(2)免疫荧光方法检测23例急性白血病患者ABCG2蛋白表达强度,从低到高依次为:正常对照组、初发组和复发组,三者之间差异有统计学意义(P0.0001)(3)激光共聚焦显微镜观察和Western-blot结果证明转染成功;(4)ABCG2过表达降低细胞的增殖能力;(5)ABCG2过表达细胞对砷剂耐受性下降。结论: (1)抗砷细胞内ABCG2蛋白表达下调,细胞抗砷性与ABCG2表达呈负相关;(2)ABCG2高表达是急性白血病患者不利的预后因素;(3)成功建立过表达ABCG2基因的细胞模型;
[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.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R346

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