龙胆苦苷通过调控miR-34c-5p/XBP1轴抑制作胃癌细胞HGC-27增殖、迁移和侵袭的分子机制研究

Gentiopicroside inhibits proliferation,migration,and invasion of gastric cancer HGC-27 cells by regulating miR-34c-5p/XBP1 axis

背景龙胆苦苷(gentiopicroside,GPS)属于环烯醚萜苷类单体化合物,可抑制肺癌和卵巢癌等肿瘤细胞的恶性行为,但其能否抑制胃癌细胞恶性行为还未知.本研究假设GPS能够通过调控微小RNA-34c-5p(microRNA-34c-5p,miR-34c-5p)/X盒结合蛋白1(X-box binding protein-1,XBP1)轴抑制胃癌细胞恶性行为.目的探讨GPS对胃癌细胞HGC-27增殖、迁移和侵袭的影响及可能机制.方法体外培养HGC-27细胞,分为对照组、不同剂量(6、30、150μg/mL)GPS组、miR-34c-5p组、miR-NC组、150μg/mL GPS+anti-miR-34c-5p组和150μg/mL GPS+anti-miR-NC组,细胞计数试剂盒-8(cell counting kit-8,CCK-8)法检测细胞增殖,Transwell检测细胞迁移和侵袭,实时定量聚合酶链式反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测细胞中miR-34c-5p表达,蛋白质印迹法检测细胞中XBP1蛋白表达.双荧光素酶报告基因实验验证miR-34c-5p和XBP1靶向调控关系.结果与对照组比较,不同剂量(6、30、150μg/mL)GPS组HGC-27细胞活性、迁移数和侵袭数及细胞中XBP1蛋白表达均降低(P<0.05),miR-34c-5p表达升高(P<0.05).miR-34c-5p组HGC-27细胞活性、迁移数和侵袭数及细胞中XBP1蛋白表达均低于对照组和miR-NC组(P<0.05).miR-34c-5p靶向负调控XBP1.与150μg/mL GPS组或150μg/mL GPS+anti-miR-NC组比较,150μg/mL GPS+anti-miR-34c-5p组HGC-27细胞活性、迁移数和侵袭数及细胞中XBP1蛋白表达均升高(P<0.05).结论龙胆苦苷可抑制胃癌细胞HGC-27增殖、迁移和侵袭,其作用机制可能调控miR-34c-5p/XBP1轴有关.

BACKGROUND Gentiopicroside(GPS)is a monomer compound of iridoid glycoside,which can inhibit the malignant behavior of lung cancer and ovarian cancer cells,but it is unknown whether it can inhibit the malignant behavior of gastric cancer cells.We hypothesized that GPS could inhibit the malignant behavior of gastric cancer cells by regulating the microRNA-34c-5p(miR-34c-5p)/X-box binding protein-1(XBP1)axis.AIM To explore the effect of GPS on the proliferation,migration,and invasion of gastric cancer cells HGC-27 and the possible mechanism.METHODS HGC-27 cells were cultured in vitro and divided into a control group,low-,medium-,and high-dose GPS groups(6,30,and 150μg/mL,respectively),a miR-34c-5p group,a miR-NC group,a 150μg/mL GPS+anti-miR-34c-5p group,and a 150μg/mL GPS+anti-miR-NC group.Cell counting kit-8(CCK-8)method was used to detect cell proliferation;cell migration and invasion were detected by Transwell assay;real-time quantitative polymerase chain reaction(RT-PCR)was used to detect the expression of miR-34c-5p.The expression of XBP1 protein in cells was detected by Western blot.The dual luciferase reporter gene assay was used to verify the targeted relationship between miR-34c-5p and XBP1.RESULTS Compared with the control group,the cell viability,the number of migrated cells,the number of invaded cells,and the expression of XBP1 protein in the GPS groups(6,30,and 150μg/mL)were significantly reduced(P<0.05),but the expression of miR-34c-5p was significantly increased(P<0.05).The cell viability,the number of migrated cells,the number of invaded cells,and the expression of XBP1 protein in the miR-34c-5p group were significantly lower than those in the control group and miR-NC group(P<0.05).miR-34c-5p could regulate the expression of XBP1.Compared with the 150μg/mL GPS group or 150μg/mL GPS+anti-miR-NC group,the viability of HGC-27 cells,the number of migrated cells,the number of invaded cells,and the expression of XBP1 protein in the 150μg/mL GPS+anti-miR-34c-5p group were significantly increased(P<0.05).CONCLUSION Gentiopicroside inhibits the proliferation,migration,and invasion of gastric cancer cells via mechanisms that may be related to the regulation of the miR-34c-5p/XBP1 axis.