干扰YAP1对食管癌Eca-109细胞生物学功能的影响

发布时间：2018-03-02 02:35

本文关键词： Eca-109细胞 YAP1 RNAi P53基因　出处：《重庆医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：应用RNAi技术研究Yes相关蛋白1(Yes-associated protein1, YAP1)对Eca-109细胞生物学功能的影响,为进一步研究YAP1在食管癌发生、发展中的作用以及食管癌基因靶向治疗方向提供一定的理论基础。方法：由上海吉凯基因化学技术有限公司构建包含针对YAPI基因的shRNA慢病毒载体,将食管癌Eca-109细胞分为三组(慢病毒转染组、空病毒转染组、空白对照组),再用慢病毒载体、空病毒载体分别转染至食管癌Eca-109细胞,采用qPCR和蛋白印迹法检测转染前后食管癌Eca-109细胞中YAPI mRNA和蛋白以及P53蛋白的表达情况；流式细胞仪检测细胞周期、凋亡情况；CCK8实验检测细胞的增殖情况变化。并对以上结果通过SPSS20.0统计软件进行统计分析。结果：慢病毒转染组细胞内YAP1 mRNA和蛋白表达量低于空白对照组和空病毒转染组,P53基因表达量高于空白对照组和空病毒转染组；Eca-109细胞转染组增值率从第3天开始低于空病毒转染组及空白对照组(P0.05)；慢病毒转染组细胞G1期比例较空白对照组和空病毒转染组增高(P0-05),早期凋亡率明显增加(P0.05),以上差距均有统计学意义。结论：结果显示干扰YAP1可成功诱导Eca-109细胞凋亡,降低Eca-109细胞的增殖能力,且YAP1抗凋亡及降低食管癌Eca-109细胞增殖能力的作用可能部分与P53基因相关。干扰YAP1可能成为食管癌基因靶向治疗的进一步研究方向。
[Abstract]:Objective: to study the effect of Yes associated protein 1- Yes-associated protein 1 (YAP1) on the biological function of Eca-109 cells by RNAi technique. Methods: the shRNA lentivirus vector containing YAPI gene was constructed by Shanghai Qikai Gene Chemical Technology Co., Ltd. Esophageal carcinoma Eca-109 cells were divided into three groups: lentivirus transfection group, empty virus transfection group and blank control group. QPCR and Western blot were used to detect the expression of YAPI mRNA, protein and p53 protein in esophageal carcinoma Eca-109 cells before and after transfection, and flow cytometry was used to detect the cell cycle. The changes of cell proliferation were detected by CCK8 assay. The results were statistically analyzed by SPSS20.0 software. Results: the expression of YAP1 mRNA and protein in lentivirus transfected group was lower than that in blank control group and empty virus group. The expression of p53 gene in the transfected group was higher than that in the blank control group and the empty virus transfection group, the proliferation rate of the transfected group was lower than that of the empty virus transfection group and the blank control group from the 3rd day, and the ratio of G1 phase of the lentivirus transfection group was higher than that of the blank control group. The rate of early apoptosis was significantly increased in both groups. Conclusion: interference of YAP1 can induce apoptosis of Eca-109 cells successfully. The inhibition of proliferation of Eca-109 cells and the role of YAP1 in inhibiting apoptosis and reducing proliferation of esophageal cancer Eca-109 cells may be partly related to p53 gene. Interfering with YAP1 may be a further research direction for gene targeting therapy of esophageal cancer.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1

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