ADPGK-AS1调控miR-1301-3p基因表达影响胃癌细胞增殖和凋亡的机制研究  

Effects and action mechanism of ADPGK-AS1 on the proliferation and apoptosis of gastric cancer cells by regulating the expression of miR-1301-3p gene in vitro

作　　者：周鉴基 王山 王太洪[2] ZHOU Jianji;WANG Shan;WANG Taihong(Department of Surgery,Yancheng Second People’s Hospital,Jiangsu,Yancheng 224000,China;不详)

机构地区：[1]江苏省盐城市第二人民医院外科,224000 [2]江苏省肿瘤医院普外科

出　　处：《河北医药》2021年第9期1285-1290,共6页Hebei Medical Journal

摘　　要：目的探讨二磷酸腺苷依赖葡糖激酶反义RNA1(ADPGK-AS1)对胃癌细胞增殖和凋亡的影响及作用机制。方法培养人胃黏膜上皮正常细胞系GES-1和胃癌细胞系AGS、HGC-27和Hs-746T,实时荧光定量PCR(RT-qPCR)检测细胞中ADPGK-AS1和微小RNA-1301-3p(miR-1301-3p)表达水平。转染ADPGK-AS1小干扰RNA或miR-1301-3p模拟物至HGC-27细胞,四甲基噻唑蓝染色法(MTT)、流式细胞术和蛋白印迹(Western blot)分别检测沉默ADPGK-AS1或过表达miR-1301-3p对HGC-27细胞增殖、凋亡及增殖细胞核抗原(PCNA)、Ki-67、剪切型聚腺苷二磷酸-核糖聚合酶(Cleaved PARP)、剪切型半胱天冬酶-3(Cleaved caspases-3)、磷酸化p38(p-p38)和磷酸化p53(p-p53)蛋白表达的影响。双荧光素酶报告基因实验验证ADPGK-AS1与miR-1301-3p调控关系。结果与GES-1细胞比较,胃癌细胞AGS、HGC-27和Hs-746T中ADPGK-AS1表达水平升高(P<0.05),miR-1301-3p表达水平降低(P<0.05)。沉默ADPGK-AS1或过表达miR-1301-3p后,HGC-27细胞存活率及PCNA、Ki-67和Cleaved caspase-3蛋白表达降低(P<0.05),细胞凋亡率和Cleaved PARP蛋白表达升高(P<0.05)。沉默ADPGK-AS1还可促进HGC-27细胞p38丝裂素活化蛋白激酶(p38MAPK)信号途径蛋白p-p38和p-p53表达(P<0.05)。ADPGK-AS1在HGC-27细胞中靶向负调控miR-1301-3p表达。抑制miR-1301-3p表达降低了沉默ADPGK-AS1对HGC-27细胞增殖、凋亡及PCNA、Ki-67、Cleaved caspase-3、Cleaved PARP、p-p38和p-p53表达的影响。结论沉默ADPGK-AS1表达抑制胃癌细胞增殖,并促进细胞凋亡,其机制与靶向上调miR-1301-3p表达和激活p38MAPK信号通路有关。Objective To investigate the effects and action mechanism of adenosine diphosphate dependent glucokinase antisense RNA-1(ADPGK-AS1)on the proliferation and apoptosis of gastric cancer cells by regulating the expression of miR-1301-3p gene in vitro.Methods Human gastric mucosal normal epithelial cell line GES 1 and gastric cancer cell lines AGS,HGC-27 and Hs-746T were cultured in vitro.The expression levels of ADPGK-AS1 and microRNA-1301-3p(miR-1301-3p)were detected by Real time quantitative PCR(RT-qPCR).After ADPGK-AS1 interfering RNA or miR-1301-3p mimics were transfected into HGC-27 cells,the effects of silencing ADPGK-AS1 or over expressing miR-1301-3p on HGC-27 cell proliferation,apoptosis and the expression of proliferating cell nuclear antigen(PCNA),Ki-67,cleavage polyadenosine diphosphate ribose polymerase(Cleaved-PARP),cleavage-caspase-3(Cleaved-caspases-3),phosphorylated p38(p-p38)and phosphorylated p53(p-p53)were detected by methyl thiazolyl tetrazolium blue staining(MTT),flow cytometry,and Western Blot,respectively.Moreover the double luciferase reporter gene test was used to verify the regulatory relationship between ADPGK AS1 and miR-1301-3p.Results As compared with those in GES-1 cells,the expression levels of ADPGK AS1 in gastric cancer cells AGS,HGC-27 and Hs-746T were significantly increased(P<0.05),however the expression levels of miR-1301-3p were significantly decreased(P<0.05).After silencing ADPGK AS1 or overexpression of miR-1301-3p,the survival rate of HGC-27 cells and the expression levels of PCNA,Ki-67,and Cleaved caspase-3 protein were significantly decreased(P<0.05),however the apoptosis rate and expression levels of Cleaved PARP were significantly increased(P<0.05).Moreover silencing ADPGK-AS1 could promote the expressions levels of p p38 and p p53(P<0.05).ADPGK-AS1 targetedly negatively regulated the miR-1301-3p expressions in HGC-27 cells.And to inhibit the miR-1301-3p expression could decrease the effects of silencing ADPGK-AS1 on the proliferation,apoptosis,and the expressions of P

关 键 词：ADPGK-AS1 miR-1301-3p 胃肿瘤 细胞增殖 凋亡 P38MAPK信号通路 

分 类 号：R735.2[医药卫生—肿瘤]