TCF3对非小细胞肺癌A549细胞株生长和迁移的影响

Effects of TCF3 on cell proliferation and migration in non-small cell cancer A549 cells

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mRNA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mRNA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 cell factor 3（TCF3） gene silencing on the Wnt pathway transcriptional activity and cell growth and migration in non-small cell cancer A549 cells. Methods：The TCF3 gene was silenced by RNAi technology and used as the experimental group，meanwhile the blank control group and negative control group were set up. The mRNA and protein expression levels were detected by real-time fluorogenic quantitative PCR and Western blot，separately. A TOPflash/FOPflash repoter gene was employed for analysis of the Wnt pathway transcriptional activity. The cell proliferation was detected by CCK-8 assay. The apoptosis and cell cycle were analyzed by flow cytometry. Transwell chamber assay was used to examine the migration potential of A549 cells. Results：Compared with those of blank and negative control groups，mRNA and protein levels of TCF3 were lower in the experimental group. The ratio of TOPflash/FOPflash was lower（P=0.00，P=0.00）. The protein expression of Wnt target C-myc was declined（P=0.00，P=0.00）. The cell proliferation was inhibited. The apoptosis was promoted and the percentage of A549 cells in G0/G1 phase was in－creased（P=0.01，P=0.00）and that in S phase was decreased（P=0.01，P=0.00）. The cell migration ability was reduced（P=0.01，P=0.01）. Conclusion：TCF3 seems to act as a transcriptional activator and promote the malignant biological in lung cancer cells.