下调lncRNA GNAS-AS1对胃癌细胞AGS增殖、侵袭、迁移及顺铂敏感性的影响

Effects of lncRNA GNAS-AS1 knockdown on the proliferation,invasion,migration and cisplatin sensitivity of the gastric cancer cells AGS

目的 探讨下调长链非编码RNA(lncRNA) GNAS-AS1对胃癌细胞AGS增殖、侵袭、迁移与顺铂敏感性的影响。方法 qRT-PCR测定胃癌细胞中GNAS-AS1表达变化。在胃癌细胞AGS中转染GNAS-AS1 siRNA,用顺铂处理,CCK-8法测定细胞增殖,平板克隆实验测定细胞克隆能力,采用流式细胞术测定凋亡,Transwell小室测定细胞迁移和侵袭,Western blot测定Cleaved Caspase-3、MMP-2、MMP-9蛋白表达。通过starbase软件分析,GNAS-AS1的靶基因,双荧光素酶报告基因实验证实二者靶向关系。在胃癌细胞AGS中共转染GNAS-AS1 siRNA、miR-362 inhibitor,同样利用上述方法测定细胞增殖、克隆、凋亡、迁移、侵袭变化。结果 GNAS-AS1在胃癌细胞中高表达。GNAS-AS1 siRNA协同顺铂抑制胃癌细胞AGS增殖、克隆、迁移、侵袭,诱导细胞凋亡。下调GNAS-AS1靶向促进miR-362表达。miR-362 inhibitor逆转下调GNAS-AS1对顺铂作用的胃癌细胞AGS增殖、克隆、凋亡、迁移、侵袭影响。结论 下调lncRNA GNAS-AS1通过调控miR-362表达,从而降低AGS的增殖、侵袭和迁移能力,增加顺铂敏感性。

Objective To investigate the effect of long non-coding RNA(lncRNA)GNAS-AS1 knockdown on the proliferation,invasion,migration,and cisplatin sensitivity of the gastric cancer cell line AGS.Methods Expression levels of GNAS-AS1 in gastric cancer cells were detected by quantitative reverse-transcription polymerase chain reaction(qRT-PCR).After transfection of GNAS-AS1 siRNA in AGS cells and induced with cisplatin,cell proliferation,cloning ability,and apoptosis were examined by cell counting kit-8(CCK-8)assay,colony formation assay and flow cytometry,respectively.Cell migration and invasion were detected by Transwell assay.Protein levels of cleaved caspase-3,matrix metalloproteinase-2(MMP-2)and MMP-9 were detected by Western blot.The target relationship between GNAS-AS1 and miR-362 was verified by Starbase prediction and dual-luciferase reporter assay.After co-transfection of GNAS-AS1 siRNA and miR-362 inhibitor,cell behaviors were similarly examined.Results GNAS-AS1 was upregulated in gastric cancer cells.Transfection of GNAS-AS1 siRNA in cisplatin-induced AGS cells inhibited the proliferation,cloning,migration,and invasion,and induced apoptosis.Knockdown of GNAS-AS1 targeted and upregulated miR-362.Transfection of miR-362 inhibitor reversed the role of GNAS-AS1 knockdown on the proliferation,cloning,apoptosis,migration and invasion of AGS cells.Conclusion Knockdown of lncRNA GNAS--S1 regulates the expression of miR-362,thereby reducing the proliferation,invasion and migration abilities of AGS and increasing cisplatin sensitivity.