摘要
目的:探讨类泛素蛋白FAT10的异常表达在肺鳞癌进展中的作用及机制。方法:采用RT-PCR和Westerm blotting法检测40例肺鳞癌及癌旁组织中FAT10 mRNA和蛋白表达水平。通过电穿孔法分别将FAT10 siRNA和重组表达载体pcDNA3.1-FAT10转染人肺鳞癌NCI-H226细胞,转染48 h后,CCK-8检测细胞增殖,流式细胞术检测细胞周期和细胞凋亡的变化,小室侵袭实验检测细胞的侵袭能力变化,RT-PCR和Westerm blotting检测Hsp90/Akt信号通路的变化。结果:FAT10基因和蛋白在人肺鳞癌组织中的表达量分别是(0.89±0.11)、(1.56±0.09),高于癌旁组织(0.42±0.06)、(0.67±0.04),差异有统计学意义(P<0.05);干扰NCI-H226细胞中FAT10表达后,NCI-H226细胞增殖(0.527±0.011)被抑制,对细胞周期分布无明显影响,细胞凋亡率(6.13±0.37)%增加,细胞的侵袭能力降低,Hsp90/Akt信号通路表达(0.25±0.04)下调;增强NCI-H226细胞中FAT10表达后,增殖能力(0.948±0.016)明显增强,对细胞周期分布无明显影响,细胞凋亡率(2.47±0.25)%降低,细胞的侵袭能力降低,Hsp90/Akt信号通路表达(0.84±0.06)上调。结论:FAT10可通过上调Hsp90/Akt信号通路表达,促进肺鳞癌细胞增殖,减少细胞凋亡,增强肺鳞癌细胞的侵袭能力,FAT10有可能成为预防和治疗作用的靶点。
Objective To explore the role and mechanism of abnormal expression of FAT10in lung squamous carcinoma.Methods The mRNA and protein levels of FAT10in40cases of lung squamous carcinoma and adjacent tissues were detected by RT-PCR and Western blotting.Transfection of FAT10siRNA and recombinant expression vector pcDNA3.1-FAT10was performed into human lung squamous cell carcinoma NCI-H226cells by electroporation.After48h of transfection,CCK-8was used to detect the cell proliferation,flow cytometry was used to detect the cell cycle and apoptosis,cell invasiveness was measured by cell invasion assay,and Hsp90/Akt signaling pathway was detected by RT-PCR and Western blotting.Results The expression levels of FAT10gene and protein in human lung squamous cell carcinoma tissues were(0.89±0.11)and(1.56±0.09),higher than those in adjacent tissues[(0.42±0.06)and(0.67±0.04)],respectively(P<0.05).After interfering with FAT10expression in NCI-H226cells,the proliferation of NCI-H226cells(0.527±0.011)was inhibited and the distribution of cell cycle was not affected.The apoptotic rate of NCI-H226cells increased significantly[(6.13±0.37)%],the invasive ability of cells decreased,and the expression of Hsp90/Akt signaling pathway decreased(0.25±0.04).After the expression of FAT10in NCI-H226cells increased,the proliferation ability increased significantly(0.948±0.016).There was no significant effect on the distribution of cell cycle,and the apoptotic rate decreased[(2.47±0.25)%].The expression of Hsp90/Akt signaling pathway was upregulated(0.84±0.06).Conclusion FAT10can up-regulate the expression of Hsp90/Akt signaling pathway,promote the proliferation of lung squamous cell carcinoma cells,reduce cell apoptosis,and enhance the invasive ability of lung squamous cell carcinoma cells.FAT10may become a target of prevention and treatment.
作者
郝术安
王钧
HAO Shu-an;WANG Jun(Tianjin Jinnan Xianshuigu Hospital, Tianjin (300350), China)
出处
《中国中西医结合外科杂志》
CAS
2018年第6期742-748,共7页
Chinese Journal of Surgery of Integrated Traditional and Western Medicine
关键词
肺鳞癌
泛素蛋白
电转法
细胞增殖
凋亡
Lung squamous carcinoma
FAT10
electroporation method
cell proliferation
apoptosis
