胃癌对奥沙利铂耐药性与miR-145关系初探

发布时间：2018-03-29 20:43

本文选题：胃癌　切入点：奥沙利铂　出处：《首都医科大学》2016年硕士论文

【摘要】：研究目的：本研究旨在探索miR-145与胃癌奥沙利铂耐药的关系。材料与方法：通过浓度递增方法诱导出对奥沙利铂耐药的胃癌细胞株BGC-823/L-OHP,应用MTT法检测两组ICso验证耐药株诱导成功。通过实时定量PCR方法检验野生BGC-823及对奥沙利铂耐药株BGC-823/L-OHP两组细胞的miR-145表达水平,验证miR-145与胃癌细胞奥沙利铂耐药相关性。构建差异表达miR-145的四组BGC-823细胞系(高表达组、低表达组、空载体组、野生组),通过MTT法检测四组细胞对奥沙利铂的半数抑制浓度(IC50),在细胞水平进一步验证miR-145与胃癌细胞奥沙利铂耐药相关性。结果：构建耐药株后,通过MTT实验显示BGC-823细胞株的半抑制浓度(IC50)为8.164ug/ml, BGC-823/L-OHP细胞株的ICso为138.994ug/ml,较前者明显升高,耐药株建立成功。通过病毒向野生BGC-823细胞系中转染靶基因,荧光显微镜下可观察到四组细胞内可见荧光蛋白,成功构建四组差异表达miR-145的胃癌细胞系(高表达组、低表达组、空载体组、野生组),MTT实验结果显示高表达miR-145组IC50为9.861ug/ml,低表达miR-145组ICso为5.377ug/ml,空载体组ICso为5.318ug/ml,野生组ICso为4.805ug/ml,高表达组表现出奥沙利铂耐药性升高。结论：miR-145在BGC-823细胞株与BGC-823/L-OHP田胞株之间表达存在差异,耐药株中表达明显升高；高表达miR-145后,胃癌细胞系BGC-823对奥沙利铂耐药性升高。miR-145可作为预测胃癌奥沙利铂化疗是否敏感的标志物,高表达miR-145的胃癌细胞奥沙利铂化疗敏感性欠佳。
[Abstract]:Objective: to explore the relationship between miR-145 and oxaliplatin resistance in gastric cancer. Materials and methods: to induce oxaliplatin resistant gastric cancer cell line BGC-823 / L-OHPby increasing concentration, and to detect the drug resistance of two groups by MTT assay. The expression of miR-145 was detected by real-time quantitative PCR assay in both groups of BGC-823 and BGC-823/L-OHP cells, which were resistant to oxaliplatin. To verify the correlation between miR-145 and oxaliplatin resistance in gastric cancer cells, four groups of BGC-823 cell lines (high expression group, low expression group, empty vector group) with differentially expressed miR-145 were constructed. In wild group, the half inhibitory concentration of oxaliplatin on oxaliplatin was detected by MTT method, and the correlation between miR-145 and oxaliplatin resistance in gastric cancer cell line was further verified at the cell level. MTT assay showed that the semi-inhibitory concentration of BGC-823 cell line IC50 was 8.164ugr / ml, and the ICso of BGC-823/L-OHP cell line was 138.994ugr / ml, which was significantly higher than that of the former cell line, and the drug resistant strain was successfully established. The target gene was transfected into the wild BGC-823 cell line by virus transfection. Four groups of gastric cancer cell lines with differentially expressed miR-145 (high expression group, low expression group, empty vector group) were successfully constructed. The results of wild group showed that the IC50 of high expression miR-145 group was 9.861 ugr / ml, the ICso of low expression miR-145 group was 5.377ugr / ml, the ICso of empty vector group was 5.318ugr / ml, the ICso of wild group was 4.805ugr / ml, and the drug resistance of oxaliplatin was increased in high expression group. There are differences in expression between two groups. After high expression of miR-145, the drug resistance of gastric cancer cell line BGC-823 to oxaliplatin was increased. MiR-145 could be used as a marker to predict the chemosensitivity of oxaliplatin to oxaliplatin in gastric cancer, but the chemosensitivity of gastric cancer cell line with high expression of miR-145 to oxaliplatin was not good.
【学位授予单位】：首都医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.2

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