S100A7蛋白在体外培养人视网膜色素上皮细胞中的表达

发布时间：2018-07-06 10:37

本文选题：细胞培养 + ARPE-19细胞　；参考：《重庆医科大学》2011年硕士论文

【摘要】：目的1掌握人视网膜色素上皮细胞系ARPE-19细胞培养、传代和冻存技术。2利用免疫荧光技术探讨S100A7蛋白在体外培养人视网膜色素上皮细胞(retinal pigment epithelial cell, RPE)中的表达及其分布状况,并联合应用Western blot技术检测S100A7蛋白在该细胞的表达。3初步探讨S100A7在ARPE-19细胞增殖中的作用,为相关疾病的诊断治疗提供新的思路。方法1利用细胞培养方法传代培养ARPE-19细胞。2取第3～6代ARPE-19细胞,运用免疫荧光技术检测S100A7蛋白在ARPE-19细胞中的表达及其分布情况。3提取ARPE-19总蛋白,运用Western blot技术检测S100A7蛋白在ARPE-19细胞的表达。4将ARPE-19细胞接种在96孔细胞培养板中,用不同浓度S100A7蛋白抗体处理细胞,然后运用MTT方法检测ARPE-19细胞增殖情况,进而初步确定S100A7在ARPE-19细胞增殖中的作用。结果1成功培养ARPE-19细胞,并且细胞能够完全满足该课题中所有实验的需要。2免疫荧光方法显示实验组中ARPE-19细胞存在S100A7蛋白的表达,表达部位为该细胞的细胞膜,而在对照组中没有该蛋白表达。3 Western blot技术进一步证实S100A7蛋白在ARPE-19细胞中的表达。4.MTT实验初步确定S100A7蛋白能够促进ARPE-19细胞的增殖过程。结论本实验首次发现并证实S100A7蛋白在人RPE细胞中的表达,并且初步确定其在RPE细胞增殖过程中起促进作用,从而为研究人视网膜疾病特别是增殖性玻璃体视网膜疾病、糖尿病视网膜疾病提供了新的方向,为该类疾病的治疗提供了新的靶点。
[Abstract]:Objective 1 to investigate the expression and distribution of S100A7 protein in human retinal pigment epithelial cell line ARPE-19 by immunofluorescence technique. The expression of S100A7 protein in ARPE-19 cells was detected by Western blot, and the role of S100A7 protein in the proliferation of ARPE-19 cells was preliminarily studied, which provided a new idea for the diagnosis and treatment of related diseases. Methods 1 ARPE-19 cells were subcultured by cell culture method. ARPE-19 cells were collected from ARPE-19 cells. The expression and distribution of S100A7 protein in ARPE-19 cells were detected by immunofluorescence technique, and the total ARPE-19 protein was extracted from ARPE-19 cells. Western blot was used to detect the expression of S100A7 protein in ARPE-19 cells. ARPE-19 cells were inoculated into 96-well cell culture plates. The cells were treated with different concentrations of S100A7 protein antibody, and then the proliferation of ARPE-19 cells was detected by MTT assay. Furthermore, the role of S100A7 in the proliferation of ARPE-19 cells was preliminarily determined. Results (1) ARPE-19 cells were successfully cultured, and the cells could fully meet the needs of all the experiments in this study. 2 immunofluorescence method showed that there was S100A7 protein expression in ARPE-19 cells in the experimental group, and the expression location was the cell membrane of ARPE-19 cells. However, the expression of S100A7 protein in ARPE-19 cells was not further confirmed by Western blot. 4. The results showed that S100A7 protein could promote the proliferation of ARPE-19 cells. Conclusion the expression of S100A7 protein in human RPE cells was found and confirmed for the first time, and it was preliminarily confirmed that S100A7 protein plays a role in promoting the proliferation of RPE cells, which may be useful for the study of human retinal diseases, especially proliferative vitreoretinopathy. Diabetic retinopathy provides a new direction and a new target for the treatment of diabetic retinopathy.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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