膀胱癌相关转录物1在胃癌组织中的表达及其对胃癌细胞增殖和自噬水平的影响

Expression of bladder cancer associated transcript-1 in gastric cancer and its regulation of proliferation and autophagy in gastric cancer cells

目的研究长链非编码RNA(lncRNA)膀胱癌相关转录物1(BLACAT1)在胃癌组织中的表达及其对胃癌细胞增殖和自噬的影响。方法实时定量PCR检测BLACAT1在胃癌组织及胃癌细胞中的表达情况;CCK-8分析敲除BLACAT1对胃癌细胞增殖的影响;免疫荧光实验用于分析敲除BLACAT1和过表达BLACAT1对胃癌细胞自噬的影响;Western blot分析敲除BLACAT1和过表达BLACAT1对胃癌细胞自噬的影响以及自噬相关蛋白ATG3的表达变化;利用共转染实验验证BLACAT1通过上调ATG3影响胃癌细胞自噬水平。结果BLACAT1在胃癌组织中和细胞系中表达均上调。敲除BLACAT1后,胃癌细胞增殖率降低,自噬水平受到抑制,ATG3表达降低。过表达BLACAT1后,胃癌细胞自噬水平增加。共转染BLACAT1质粒和si-ATG3后,胃癌自噬水平受到抑制。结论LncRNA BLACAT1能够促进胃癌细胞的增殖和自噬,并且BLACAT1对胃癌细胞自噬水平的影响可能通过调控ATG3基因实现的。

Objective To investigate the expression of long non-coding RNA(lncRNA)bladder cancer associated transcript-1(BLACAT1)in the gastric cancer and effect of lncRNA BLACAT1 on cell proliferation and autophagy in gastric cancer cells.Methods Quantitative Real-time PCR was used to measure the expression of lncRNA BLACAT1 in gastric cancer tissues and gastric cancer cells.The proliferation levels of gastric cancer cells was analyzed by CCK-8.Immunofluorescence was used to analyze autophagy activity.Western blot was used to analyze the effect of knockdown lncRNA BLACAT1 on autophagy and ATG3 expression.The co-transfection experiment was used to verify that BLACAT1 affects autophagy level by upregulation of ATG3.Results The expression level of lncRNA BLACAT1 was up-regulated in the gastric cancer tissues than in the adjacent tissues.Knockdown lncRNA BLACAT1 could limit cells proliferation and autophagy of gastric cancer cells,and ATG3 expression was decreased.After co-transfection of pcDNA3.1-BLACAT1 and si-ATG3 into gastric cancer cells,autophagy was inhibited in gastric cancer cell lines.Conclusion LncRNA BLACAT1 could promote proliferation and autophagy in the gastric cancer cells,the promotion of lncRNA BLACAT1 on autophagy might be related to ATG3,which is expected to be a new therapeutic target for gastric cancer.