沉默FAT10基因抑制人胆囊癌GBC-SD细胞增殖、侵袭及迁移

Effect of FAT10 gene silencing on proliferation and migration of human gallbladder cancer cells GBC-SD

目的探究电转法沉默FAT10基因的表达对人胆囊癌GBC-SD细胞侵袭、增殖及迁移的影响.方法设计及合成靶向FAT10的siRNA序列(FAT10-siRNA)以及阴性对照序列(NC-siRNA),电转法转染至人胆囊癌GBC-SD细胞,分别形成FAT10-GBC-SD、NC-siRNA-GBC-SD细胞.通过RT-PCR、Western blotting检测电转后细胞中FAT10 mRNA与蛋白的表达情况,流式细胞术检验细胞凋亡及周期,迁移,细胞侵袭实验分别测定细胞增殖,划痕愈合实验,CCK-8增殖实验,以及侵袭的能力.结果FAT10-GBC-SD细胞中FAT10 mRNA和蛋白表达较NC-siRNA-GBC-SD细胞和空载组(Ctrl-GBC-SD细胞组)显著降低.FAT10-GBC-SD细胞的成长速度显著减慢,细胞周期阻止在G0/G1期,S期细胞数减少;与空载组相比,FAT10-GBC-SD细胞的凋亡率显然升高(P<0.05),迁移、增殖、侵袭能力明显下降(P<0.05).结论电转法沉默FAT10基因的表达可抑制胆囊癌细胞的迁移、侵袭及增殖能力.

Objective To investigate the effects of the expression of silent FAT10 gene by electroconversion on the invasion,proliferation and migration of GBC-SD cells from human gallbladder carcinoma.Methods SiRNA sequences(FAT10-siRNA)and negative control sequences(NC-siRNAs)targeting FAT10 were designed and synthesized.Electrochemical transfer was transmitted to human gallbladder carcinoma GBC-SD cells.FAT10-GBC-SD and NC-siRNA-GBC-SD cells are formed respectively.And...The expression of FAT10 mRNA and protein in cells after electroconversion was detected by RT-PCR and Western blot,and apoptosis and cycle were detected by flow cytometry,migration,and cell invasion experiments.Cell proliferation,scratch healing experiments,CCK-8 proliferation experiments,And the ability to invade.Results FAT10-GBC-SD cells showed significantly lower expression of FAT10 mRNA and protein than NC-siRNA-GBC-SD cells and empty carriers(Ctrl-GBC-SD cells).The growth rate of FAT10-GBC-SD cells slowed significantly,and the cell cycle prevented the reduction of the number of S cells in the G0/G1 phase;Compared with the empty load group,the apoptosis rate of FAT10-GBC-SD cells was significantly higher(P&It;0.05),the ability to migrate,proliferate,and invade is significantly reduced(P&It;0.05).Conclusion The expression of the silent FAT10 gene can inhibit the migration,invasion and proliferation of gallbladder cancer cells.