热休克蛋白90α对食管癌细胞侵袭转移能力的影响
发布时间：2018-08-09 18:55
【摘要】：目的：探讨HSP90α在食管癌细胞侵袭转移中的作用机制。方法：使用细胞转染技术将siRNA-HSP90α转染入CE81T-4细胞中,并分为实验组、阴性对照组、空白对照组;并用RT-PCR和Western blot检测3组细胞中HSP90αmRNA水平和蛋白质水平的表达差异,验证是否转染成功。MTT法检测3组细胞的增值活力,流式细胞术分析细胞周期、凋亡的改变,划痕实验和Transwell体外侵袭实验检测转染后CE81T-4细胞迁移和侵袭的变化。结果：在siRNA-HSP90α成功转染CE81T-4细胞后,RT-PCR检测3组结果显示,实验组中HSP90αmRNA水平表达量显著低于空白对照组和阴性对照组(P 0.05, P 0.05)。Western blot分析显示实验组HSP90α蛋白表达量降低,与空白对照组和阴性组相比差异有统计学意义(P0.05,P0.05)。MTT实验结果显示,实验组细胞的增殖能力明显减弱,与其他两组比较差异具有统计学意义(P0.05,P0.05)。HSP90a基因表达沉默后,细胞凋亡率为32.67%,较空白对照组和阴性对照组明显增加(P0.05,P0.05)。实验组细胞周期被阻滞在G0/G1期。Transwell体外侵袭实验显示实验组侵袭迁移的细胞个数为(76.4±8.7),显著低于空白照组(140.3±5.8,P0.05)与阴性对照组(129.3±10.1,P0.05)。划痕实验结果显示,72h后实验组细胞的迁移力较其他两组明显降低。结论：HSP90a的下调会降低食管癌细胞的转移侵袭能力。
[Abstract]:Objective: to investigate the role of HSP90 伪 in invasion and metastasis of esophageal cancer cells. Methods: siRNA-HSP90 伪 was transfected into CE81T-4 cells by cell transfection technique, and was divided into experimental group, negative control group and blank control group, and the expression of HSP90 伪 mRNA and protein in the three groups were detected by RT-PCR and Western blot. The cell cycle and apoptosis were analyzed by flow cytometry, and the changes of migration and invasion of CE81T-4 cells after transfection were detected by scratch test and Transwell invasion assay in vitro. Results: after siRNA-HSP90 伪 was successfully transfected into CE81T-4 cells, the results of RT-PCR showed that the expression of HSP90 伪 mRNA in the experimental group was significantly lower than that in the blank control group and the negative control group (P 0.05, P 0.05). Western blot analysis showed that the expression of HSP90 伪 protein in the experimental group was significantly lower than that in the control group. Compared with the blank control group and the negative group, the difference was statistically significant (P 0.05 P 0.05). MTT assay showed that the proliferation ability of the experimental group was significantly decreased, and the difference was statistically significant compared with the other two groups (P 0.05 P 0.05) .HSP90a gene expression silenced. The apoptosis rate was 32.67%, which was significantly higher than that in the blank control group and the negative control group (P 0.05). The cell cycle of the experimental group was blocked in the G0/G1 phase. Transwell invasion in vitro showed that the number of the cells in the experimental group was (76.4 卤8.7), significantly lower than that in the blank irradiation group (140.3 卤5.8) and the negative control group (129.3 卤10.1). The results of scratch test showed that the migration ability of the experimental group was significantly lower than that of the other two groups after 72 hours. Conclusion the down-regulation of% HSP90a may decrease the metastasis and invasion ability of esophageal cancer cells.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1
，
本文编号：2174990
[Abstract]:Objective: to investigate the role of HSP90 伪 in invasion and metastasis of esophageal cancer cells. Methods: siRNA-HSP90 伪 was transfected into CE81T-4 cells by cell transfection technique, and was divided into experimental group, negative control group and blank control group, and the expression of HSP90 伪 mRNA and protein in the three groups were detected by RT-PCR and Western blot. The cell cycle and apoptosis were analyzed by flow cytometry, and the changes of migration and invasion of CE81T-4 cells after transfection were detected by scratch test and Transwell invasion assay in vitro. Results: after siRNA-HSP90 伪 was successfully transfected into CE81T-4 cells, the results of RT-PCR showed that the expression of HSP90 伪 mRNA in the experimental group was significantly lower than that in the blank control group and the negative control group (P 0.05, P 0.05). Western blot analysis showed that the expression of HSP90 伪 protein in the experimental group was significantly lower than that in the control group. Compared with the blank control group and the negative group, the difference was statistically significant (P 0.05 P 0.05). MTT assay showed that the proliferation ability of the experimental group was significantly decreased, and the difference was statistically significant compared with the other two groups (P 0.05 P 0.05) .HSP90a gene expression silenced. The apoptosis rate was 32.67%, which was significantly higher than that in the blank control group and the negative control group (P 0.05). The cell cycle of the experimental group was blocked in the G0/G1 phase. Transwell invasion in vitro showed that the number of the cells in the experimental group was (76.4 卤8.7), significantly lower than that in the blank irradiation group (140.3 卤5.8) and the negative control group (129.3 卤10.1). The results of scratch test showed that the migration ability of the experimental group was significantly lower than that of the other two groups after 72 hours. Conclusion the down-regulation of% HSP90a may decrease the metastasis and invasion ability of esophageal cancer cells.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1
，
本文编号：2174990
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