TCF3对非小细胞肺癌A549细胞株生长和迁移的影响

发布时间：2018-03-06 05:28

本文选题：T细胞因子　切入点：A　出处：《重庆医科大学学报》2015年03期 　论文类型：期刊论文

【摘要】：目的:探讨T细胞因子3(T cell factor 3,TCF3)基因沉默对非小细胞肺癌A549细胞株的Wnt通路转录活性及细胞生长和迁移的影响。方法:采用RNAi技术将TCF3基因沉默作为实验组,同时设立空白对照组和阴性对照组。采用实时荧光定量PCR及Western blot检测各组细胞中TCF3的m RNA和蛋白表达水平,通过TOPFlash/FOPFlash双荧光素酶报告基因检测Wnt通路转录活性,CCK-8检测细胞增殖,Transwell检测细胞迁移能力。结果:与空白对照组和阴性对照组比较,实验组TCF3的m RNA和蛋白水平明显下调;TOPflash/FOPflash报告基因的比值降低(P=0.00,P=0.00);Wnt靶基因C-myc的蛋白表达量下降(P=0.00,P=0.00);细胞的增殖活性降低,细胞凋亡率增加(P=0.00,P=0.00),细胞周期比例发生变化,G0/G1期细胞增多(P=0.01,P=0.00),S期细胞减少(P=0.01,P=0.00);细胞迁移能力减弱(P=0.00,P=0.00)。结论:在肺癌细胞中,TCF3可能作为转录激活因子对细胞的恶性生物学行为起着正向调控作用。
[Abstract]:Objective: to investigate the effect of T cytokine 3T cell factor 3 (TCF3) gene silencing on the transcriptional activity of Wnt pathway and cell growth and migration in non-small cell lung cancer cell line A549. Methods: TCF3 gene silencing was used as experimental group by RNAi technique. At the same time, blank control group and negative control group were established. The expression of m RNA and protein of TCF3 was detected by real-time fluorescence quantitative PCR and Western blot. TOPFlash/FOPFlash double luciferase reporter gene was used to detect the transcriptional activity of Wnt pathway and CCK-8 was used to detect cell proliferation. The level of m RNA and protein of TCF3 in the experimental group decreased significantly the ratio of topp flash / FOPflash reporter gene. The protein expression of c-myc, the target gene C-myc, was decreased, and the proliferative activity of the cells was decreased. Apoptosis rate increased, cell cycle ratio changed and cell cycle ratio increased in G _ 0 / G _ 1 phase. The number of cells in the G _ 0 / G _ 1 phase increased. The number of cells decreased in the S phase, and the migration ability of the cells decreased. Conclusion: TCF3 may be a transcriptional activator in the malignant biological process of lung cancer cells, and it is possible that TCF3 may act as a transcriptional activator in the malignant biology of lung cancer cells. In order to play a positive regulatory role.
【作者单位】：重庆医科大学附属第一医院呼吸与危重症学科;
【基金】：国家自然科学基金资助项目(编号:81201841) 国家临床重点专科建设经费资助项目(编号:卫办医改函[2012]649号)
【分类号】：R734.2

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