摘要
目的探讨下调微小核糖核酸(miRNA,miR)-572对人胃癌细胞凋亡和迁移能力的影响及机制。方法实时荧光定量PCR(quantitative real-time PCR,qPCR)法检测miR-572,第10号染色体缺失的磷酸酶、张力蛋白同源物基因(phosphatase and tensin hmmlogydeleted on ten,PTEN)和蛋白激酶2(protein kinase 2,AKT2)在不同胃癌细胞株(HGC-27,AGS和SGC-7901)和正常胃黏膜上皮细胞GES-1中的表达情况。在胃癌细胞株AGS细胞中加入miR-572 inhibitor后,CCK8检测细胞活力;Tranwell实验检测细胞侵袭和迁移能力;流式细胞术检测细胞凋亡比例;qPCR检测miR-572,PTEN和AKT2的表达量。结果miR-572和AKT2在胃癌细胞系HGC-27(6.97±1.62,4.98±1.34),AGS(7.21±1.32,5.39±1.14)和SGC-7901(5.97±1.44,4.02±1.02)中较正常胃黏膜上皮细胞GES-1表达升高(1.00±0.24,1.00±0.21),PTEN表达降低(0.49±0.16,0.39±0.11,0.54±0.33 vs 1.00±0.13),差异均有统计学意义(t=2.727~3.197,均P<0.05);与对照组比较,转染miR-572 inhibitor后,miR-572和AKT2在AGS中表达下调(P<0.05),PTEN表达上调(P<0.05);CCK8实验结果显示转染miR-572 inhibitor后,与对照组比较miR-572抑制剂组细胞活力降低(P<0.05);Transwell实验发现,与对照组比较miR-572抑制剂组细胞的侵袭和迁移能力降低(P<0.05);流式细胞实验结果表明,与对照组相比miR-572抑制剂组细胞的凋亡比例降低(P<0.05)。结论下调miR-572可抑制胃癌细胞的凋亡、侵袭和迁移,其机制可能是通过PTEN/AKT2信号通路。
Objective To investigate the effect and mechanism of down-regulation of miR-572 on the apoptosis and migration of human gastric cancer cells.Methods qPCR method was used to detect the expression of miR-572,phosphatase and tensin hmmlogydeleted on ten(PTEN)and AKT2 in HGC-27,AGS and SGC-7901 gastric cancer cell lines and GES-1 normal gastric mucosal epithelial cells.After transduction of miR-572 inhibitor in AGS cells,CCK8 was used to detect cell viability.Transwell was used to detect cell invasion and migration ability.Flow cytometry was used to detect the proportion of apoptosis cells,and qPCR was used to detect the expression of miR-572,PTEN and AKT2.Results Compared with normal gastric mucosal epithelial cells GES-1,the expression of miR-572 and AKT2 increased in HGC-27(6.97±1.62,4.98±1.34),AGS(7.21±1.32,5.39±1.14)and SGC-7901(5.97±1.44,4.02±1.02)gastric cancer cell lines(1.00±0.24,1.00±0.21),while the expression of PTEN decreased(0.49±0.16,0.39±0.11,0.54±0.33 vs 1.00±0.13),the differences were statistically significant(t=2.727~3.197,all P<0.05).Compared with the control group,after transfection with miR-572 inhibitor,the expression of miR-572 and AKT2 in AGS down-regulated(P<0.05),and the expression of PTEN up-regulated(P<0.05).CCK8 experiment results showed that after transfection with miR-572 inhibitor the cell viability of the miR-572 inhibitor group reduced compared with the control group(P<0.05).Transwell experiment found that the cell invasion and migration ability of the miR-572 inhibitor group reduced compared with the control group(P<0.05),and the results of flow cytometry showed that compared with the control group,the proportion of apoptosis cell in the miR-572 inhibitor group was lower(P<0.05).Conclusion Down-regulation of miR-572 can inhibit the apoptosis,invasion and migration of gastric cancer cells,which may be explained by directly targeting the PTEN/AKT2 signaling pathway.
作者
郭殿华
程芃
陈卿奇
程正
GUO Dian-hua;CHENG Peng;CHEN Qing-qi;CHENG Zheng(Hainan Western Central Hospital,Hainan Danzhou 571756,China)
出处
《现代检验医学杂志》
CAS
2022年第3期157-161,共5页
Journal of Modern Laboratory Medicine
基金
海南省卫生医药卫生科研项目(2001320114A2001)。
关键词
胃癌
细胞侵袭
细胞凋亡
细胞增殖
细胞迁移
gastric cancer
cell invasion
cell apoptosis
cell proliferation
cell migration
