TCF4对人皮肤鳞状细胞癌A431细胞株生物学行为的影响.docVIP

PAGE PAGE 4 TCF4对人皮肤鳞状细胞癌A431细胞株生物学行为的影响 刘 燕，熊 亚，宋志强，沈 柱，郝 飞，杨希川 （400038 重庆，第三军医大学西南医院皮肤科，重庆市皮肤性病研究所） [摘要] 目的 探讨T细胞因子4(T cell factor4, TCF4)小干扰RNA（siRNA）对人皮肤鳞状细胞癌A431细胞株TCF4基因表达的抑制作用及对细胞增殖、迁移、侵袭及周期的影响。方法 转染TCF4 siRNA至A431细胞株。应用RT-PCR检测A431细胞TCF4 mRNA的表达，Western blot检测TCF4蛋白的表达，CCK-8方法检测上述各组细胞增殖率（1、2、3、4d及5d），小室迁移实验检测细胞迁移情况，Transwell方法检测侵袭情况，流式细胞仪检测细胞周期（24h）。结果 与未处理的A43l细胞株和转染空载体的A431细胞株比较，转染TCF4 siRNA表达载体1、2、3、4d及5d的细胞增殖率明显降低；TCF4 siRNA表达载体组细胞迁移和侵袭能力明显下降；细胞周期分析发现转染TCF4 siRNA使得A431细胞株出现明显的G2/M期阻滞现象。结论 采用TCF4特异性siRNA抑制皮肤鳞状细胞癌TCF4的表达，从而抑制皮肤鳞状细胞癌的增殖，为皮肤鳞状细胞癌的临床治疗提供一种新的思路。 [关键词] TCF4；鳞状细胞癌；增殖；凋亡；Wnt/β-catenin信号通路 [中图法分类号] [文献标志码] A Investigation of effect of TCF4 on the biological behavior of human squamous carcinoma cells Liu Yan, Xiong Ya, Song Zhiqiang, Shen Zhu, Hao Fei, Yang Xichuan (Department of Dermatology, Chongqing Institute of Dermatovenerology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China ) [Abstract] Objective To investigate the inhibitory effect of TCF4 siRNA on targeting TCF4 gene expression, migration, invasion and cell cycle of human squamous carcinoma cells. Methods A431 cells were transfected with TCF4 siRNA Liposome. The mRNA expression of TCF4 in A431 cells was detected by RT-PCR, the expression of TCF4 protein was detected by Western blotting. The proliferation, migration, invasion activities of A431cells and cell cycle were measured by CCK-8 method, chamber cell?migration?assay, transwell method and flow cytometry, respectively. Results The transfected TCF4 siRNA expression could inhibit significantly the expression in transcription and translation level in comparison with the un-treated A431 cells. The rates of cell proliferation, ability of migration and invasion were also significantly decreased. In addition, analysis of cell cycle demonstrated a marked blockage of the G2/M stage of the A431 cells treated with TCF4 siRNA. Conclusions It is conc