LINC00659在胃癌组织中的表达及其对人胃癌细胞侵袭、迁移和自噬的影响

目的:在胃癌组织中检测LINC00659的表达水平，并探讨LINC00659对人胃癌AGS、HGC-27细胞侵袭、迁移和自噬的影响。方法:收集宁夏回族自治区人民医院自2020年01月至11月的17例胃癌患者的肿瘤组织及癌旁组织标本，qRT-PCR法检测人胃癌组织与癌旁组织中LINC00659的表达，Western blot法和免疫组化检测人胃癌与癌旁组织中自噬相关蛋白LC3、p62的表达；在AGS和HGC-27细胞株中分别构建敲减和过表达LINC00659胃癌稳转株细胞，分为Vector组、sh-LINC00659组、pEX-3组和LINC00659组。慢病毒转染48 h后通过倒置荧光显微镜和qRT-PCR检测转染效率；Transwell、细胞划痕实验法检测各组细胞的侵袭和迁移能力；Western blot、免疫荧光检测各组细胞中自噬相关蛋白LC3、p62的相对表达水平。结果:LINC00659及自噬相关蛋白LC3在胃癌组织中的表达均显著高于癌旁组织，而自噬相关蛋白p62在胃癌组织中的表达要低于癌旁组织（均P＜0.05）。AGS、HGC-27细胞经慢病毒转染后，与空载组相比，敲减组中LINC00659相对表达水平明显降低，过表达组LINC00659相对表达水平明显升高（均P<0.01）；Transwell、细胞划痕实验结果显示，与对照组相比，AGS敲减组细胞的侵袭和迁移能力明显受到抑制，而HGC-27过表达组细胞的侵袭和迁移能力显著增强（均P＜0.05）；Western blot、免疫荧光结果显示,AGS敲减组细胞中自噬相关蛋白LC3的表达有所下调，而p62的表达则明显升高，且在HGC-27过表达组中自噬相关蛋白LC3、p62的表达水平与敲减组完全相反（均P<0.05）。结论:LINC00659在胃癌组织中高表达，且能够促进人胃癌细胞的侵袭、迁移和自噬。

Expression of LINC00659 in gastric cancer tissues and its effect on invasion,migration and autophagy of human gastric cancer cells

Objective:To detect the expression of LINC00659 in gastric cancer tissues and investigate the effects of LINC00659 on invasion,migration and autophagy of human gastric cancer cells AGS and HGC-27.Methods:The tumor tissue and paracancer tissue samples of 17 patients with gastric cancer were collected from the People's Hospital of Ningxia Hui Autonomous Region from January to November 2020.The expression of LINC00659 in human gastric cancer tissue and paracancer tissue was detected by qRT-PCR,and the expression of autophagy-asociated protein LC3 and p62 in human gastric cancer and paracancer tissue was detected by Western blot and immunohistochemistry.Knockdown and overexpression LINC00659 gastric cancer stable cells were constructed in AGS and HGC-27 cell lines,respectively,and divided into Vector group,sh-LINC00659 group,pEX-3 group and LINC00659 group.After After 48 h lentivirus transfection,transfection efficiency was detected by inverted fluorescence microscope and qRT-PCR.Transwell,scratch assay was used to detect the invasion and migration of cells in each group.Western blot,immunofluorescence was used to detect the relative expression levels of autophagy related protein LC3,p62 in each group.Results:The expression of LINC00659 and autophagy-associated protein LC3 in gastric cancer tissues was significantly higher than that in adjacent tissues,while the expression of autophagy-associated protein p62 in gastric cancer tissues was lower than that in adjacent tissues(all P<0.05).After transfection with lentivirus,the relative expression level of LINC00659 in AGS,HCC-27 cells was significantly decreased in the knockdown group and significantly increased in the overexpression group compared knockdown with the no-load group(all P<0.01).Transwell,scratch test results showed that compared with the control group,the invasion and migration of AGS knockdown cells were significantly inhibited,while the invasion and migration of HGC-27 overexpressed cells were significantly enhanced(all P<0.05).Western blot,immunofluorescence results showed that the expression of autophagy related protein LC3 was down-regulated in the AGS knockout group,while the expression of p62 was significantly increased,and the expression level of autophagy related protein LC3,p62 in the HGC-27 overexpression group was completely opposite to that in the knockout group（all P＜0.05).Conclusion:LINC00659 is highly expressed in gastric cancer tissues and can promote the invasion,migration and autophagy of human gastric cancer cells.