LINC00511靶向miR-497-5p调控胃癌细胞增殖、迁移和侵袭及机制研究

目的:探讨LINC00511对胃癌细胞增殖、迁移和侵袭的影响及其作用机制。方法:将pcDNA、pcDNA-LINC00511、si-NC、si-LINC00511、miR-NC、miR-497-5p分别转染至MGC-803细胞中，分别记为pcDNA组、pcDNA-LINC00511组、si-NC组、si-LINC00511组、miR-NC组、miR-497-5p组；将si-LINC00511质粒分别与anti-miR-NC、anti-miR-497-5p共转染至MGC-803细胞中，分别记为si-LINC00511+anti-miR-NC组、si-LINC00511+anti-miR-497-5p组。实时荧光定量PCR（RT-qPCR）检测miR-497-5p和LINC00511表达水平；蛋白质印迹（Western Blot）法检测细胞周期素D1（cyclin D1，CyclinD1）、p21、基质金属蛋白酶2（matrix metalloproteinase 2，MMP2）、基质金属蛋白酶9（matrix metalloproteinase 9，MMP9）蛋白表达水平；四甲基偶氮唑盐比色法(MTT)检测细胞活性；Transwell检测细胞迁移和侵袭；双荧光素酶报告基因实验检测LINC00511和miR-497-5p的靶向关系。结果:与正常胃黏膜上皮细胞GES-1相比，胃癌细胞MGC-803、MKN-45、AGS中miR-497-5p表达水平显著降低，LINC00511表达水平显著升高。LINC00511靶向调控miR-497-5p的表达。抑制LINC00511表达和miR-497-5p过表达可降低细胞活性和迁移、侵袭数量，降低CyclinD1、MMP2、MMP9蛋白表达水平，提高p21蛋白表达水平。干扰miR-497-5p表达逆转了抑制LINC00511表达对胃癌MGC-803细胞增殖、迁移和侵袭的抑制作用。结论:抑制LINC00511表达可抑制胃癌细胞增殖、迁移和侵袭，其机制可能与miR-497-5p表达有关，将为胃癌的治疗提供新思路和新靶点。

The mechanism of LINC00511 targeting miR-497-5p to regulate proliferation,migration and invasion of gastric cancer cells

Objective:To investigate the effect of LINC00511 on proliferation,migration and invasion of gastric cancer cells and its mechanism.Methods:pcDNA,pcDNA-LINC00511,si-NC,si-LINC00511,miR-NC,miR-497-5p were transfected into MGC-803 cells,respectively,and recorded as pcDNA group,pcDNA-LINC00511 group,si-NC group,si-LINC00511 group,miR-NC group and the miR-497-5p group.The si-LINC00511 plasmid was co-transfected into the MGC-803 cells with anti-miR-NC and anti-miR-497-5p,respectively,and recorded as si-LINC00511+anti-miR-NC group,si-LINC00511+anti-miR-497-5p group.Real-time quantitative PCR (RT-qPCR) was used to detect the expressions of miR-497-5p and LINC00511.Western Blot was used to detect cyclin D1 (CyclinD1),p21,matrix metalloproteinase 2 (MMP2),matrix metalloproteinase 9 (MMP9) expression level.Tetramethylazozolium salt colorimetric assay (MTT) was used to detect cell viability.Transwell was used to detect cell migration and invasion.Dual luciferase reporter gene experiment was used to detect targeting relationship between LINC00511 and miR-497-5p.Results:Compared with normal gastric mucosal epithelial cell GES-1,the expression of miR-497-5p in gastric cancer cells MGC-803,MKN-45 and AGS was significantly decreased,and the expression of LINC00511 was significantly increased.LINC00511 targeted the regulation of miR-497-5p expression.Inhibition of the expression of LINC00511 and overexpression of miR-497-5p can decrease the cell activity,migration and invasion,decrease the expression of CyclinD1,MMP2 and MMP9,and increase the expression of p21.Interference with miR-497-5p expression reversed the inhibitory effect of inhibition of LINC00511 expression on proliferation,migration and invasion of gastric cancer MGC-803 cells.Conclusion:Inhibition of LINC00511 expression can inhibit the proliferation,migration and invasion of gastric cancer cells.The mechanism may be related to the expression of miR-497-5p,which will provide new ideas and targets for the treatment of gastric cancer.