LncRNA LBX2-AS1调节miR-873-5p/G6PD轴对胃癌细胞增殖迁移和侵袭的影响

Effects of LncRNA LBX2-AS1 on the Proliferation Migration and Invasion of Gastric Cancer Cells by Regulating the miR-873-5p/G6PD Axis

目的:探讨长链非编码RNA(LncRNA)瓢虫同源盒2反义RNA1(LBX2-AS1)调节微小RNA-873-5p(miR-873-5p)/葡萄糖-6-磷酸脱氢酶(G6PD)轴对胃癌(GC)细胞增殖、迁移和侵袭的影响。方法:以SGC7901细胞为研究对象,将其随机分为Control组、sh-NC组、sh-LBX2-AS1组、sh-LBX2-AS1+inhibitor-NC组、sh-LBX2-AS1+miR-873-5p inhibitor组;qRT-PCR法检测LncRNA LBX2-AS1、miR-873-5p、G6PD的表达;MTT法和平板克隆实验检测SGC7901细胞增殖;划痕实验检测SGC7901细胞迁移;Transwell实验检测SGC7901细胞的侵袭;Western blot检测SGC7901细胞中MMP-2、PCNA、MMP-9、G6PD蛋白表达;荧光素酶报告基因实验检测LncRNA LBX2-AS1与miR-873-5p,miR-873-5p与G6PD之间的相互作用;小鼠荷瘤实验验证敲除LncRNALBX2-AS1对CC肿瘤生长及G6PD表达的影响。结果:sh-LBX2-AS1组SGC7901细胞中A490值、克隆数、划痕愈合率、侵袭数、LncRNA LBX2-AS1、G6PD mRNA和PCNA、MMP-2、MMP-9蛋白表达低于sh-NC组、Control组,miR-873-5p表达高于sh-NC组、Control组(P<0.05);与sh-LBX2-AS1组、sh-LBX2-AS1+inhibitor-NC组相比,sh-LBX2-AS+miR-873-5p inhibitor组miR-873-5p表达降低,A490值、克隆数、划痕愈合率、侵袭数、G6PD mRNA和PCNA、MMP-2、MMP-9蛋白表达升高(P<0.05)。LncRNA LBX2-AS1靶向负调控miR-873-5p,miR-873-5p靶向负调控G6PD。实验组移植瘤质量、体积、Ki-67阳性率、G6PD阳性率低于对照组(P<0.05)。结论:敲除LncRNA LBX2-AS1可能抑制GC细胞的增殖、迁移和侵袭,其机制可能是调节miR-873-5p/G6PD轴实现的。

Objective:To investigate the effects of long non-coding RNA(LncRNA)ladybird homeobox 2 antisense RNA1(LBX2-AS1)on the proliferation,migration,and invasion of gastric cancer(GC)cells by regulating the micro RNA-873-5p(miR-873-5p)/glucose-6-phosphate dehydrogenase(G6PD)axis.Methods:SGC7901 cells were randomly separated into control group,sh-NC group,sh-LBX2-AS1 group,sh-LBX2-AS1+inhibitor NC group,and sh-LBX2-AS1+miR-873-5p inhibitor group.qRT-PCR method was applied to detect the expression of LncRNA LBX2-AS1,miR-873-5p,and G6PD.MTT method and plate cloning experiment were applied to detect the proliferation of SGC7901 cells.Scratch experiment was applied to detect the migration of SGC7901 cells.Transwell experiment was applied to detect the invasion of SGC7901 cells.Western blot was applied to detect the expression of MMP-2,PCNA,MMP-9,and G6PD proteins in SGC7901 cells.Luciferase reporter gene assay was applied to detect the interaction between LncRNA LBX2-AS1 and miR-873-5p,and between miR-873-5p and G6PD.Mouse tumor bearing experiment was applied to verify the effects of knocking out LncRNALBX2-AS1 on CC tumor growth and G6PD expression.Results:The A490 value,clone number,scratch healing rate,invasion number,LncRNA LBX2-AS1,G6PD mRNA and PCNA,MMP-2,MMP-9 protein expression in SGC7901 cells in sh-LBX2-AS1 group were lower than those in sh-NC group and control group,while miR-873-5p expression was higher than that in sh-NC group and control group(P<0.05).Compared with the sh-LBX2-AS1 group and the sh-LBX2-AS1+inhibitor NC group,the sh-LBX2-AS+miR-873-5p inhibitor group showed a decrease in miR-873-5p expression,and an increase in A490 value,clone number,scratch healing rate,invasion number,G6PD mRNA and PCNA,MMP-2,MMP-9 protein expression(P<0.05).LncRNA LBX2-AS1 targeted the negative regulation of miR-873-5p,and miR-873-5p targeted the negative regulation of G6PD.The quality,volume,Ki-67 positive rate,and G6PD positive rate of transplanted tumors in the experimental group were lower than those in the control group(P<0.05).Conclusion:Knocking out LncRNA LBX2-AS1 may inhibit the proliferation,migration,and invasion of GC cells,and its mechanism may be achieved by regulating the miR-873-5p/G6PD axis.