微小RNA-876-3p靶向肽基脯氨酰顺反异构酶A调控胃癌细胞增殖和顺铂耐药性的作用机制研究  

Effect and mechanism of the microRNA-876-3p targets peptidyl-prolyl cis/trans isomerase A to regulates proliferation and cisplatin resistance of gastric cancer cells

作　　者：徐龙健[1] 高建超[1] 钟轩[1] 孙敬国[1] 李东坤[1] XU Long-jian;GAO Jian-chao;ZHONG Xuan;SUN Jing-guo;LI Dong-kun(Department of General Surgery,Kailuan General Hospital,Kailuan 063000,Hebei Province,China)

机构地区：[1]开滦总医院普通外科,河北开滦063000

出　　处：《中国临床药理学杂志》2020年第17期2686-2690,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究微小RNA-876-3p(miR-876-3p)通过调控肽基脯氨酰顺反异构酶A(PPIA)对胃癌细胞增殖和顺铂(DDP)耐药性的影响。方法体外培养胃癌细胞MKN28,用低浓度梯度递增方法诱导MKN28细胞形成顺铂耐药性,以构建顺铂耐药细胞株(MKN28/DDP)。MKN28和MKN28/DDP细胞各包含4个实验组:阴性对照组、第1转染组(转染si-PPIA)、第2转染组(转染miR-876-3p mimics)和第3转染组(转染si-PPIA+miR-876-3p inhibitor)。用顺铂1μg·mL^-1处理各组MKN28细胞,用顺铂5μg·mL^-1处理各组MKN28/DDP细胞,培养24 h。以实时荧光定量-PCR法检测各组细胞中miR-876-3p表达水平,蛋白质印迹法检测各组细胞中PPIA蛋白的表达水平,细胞计数试剂盒法检测各组细胞增殖活力,流式细胞术检测各组细胞细胞凋亡水平。结果miR-876-3p在正常胃上皮细胞GES-1、胃癌细胞MKN28和MKN28/DDP中的相对表达量分别为1.01±0.21,0.41±0.15和0.32±0.11,MKN28与GES-1比较,差异有统计学意义(P<0.01);MKN28/DDP与MKN28比较,差异有统计学意义(P<0.05)。在表达miR-876-3p实验中,1μg·mL^-1顺铂处理MKN28,阴性对照组和第3转染组的细胞活力分别为(1.00±0.04)%,(0.65±0.03)%;这2组的细胞凋亡率(41.87±1.65)%,(68.98±4.12)%。5μg·mL^-1顺铂处理MKN28/DDP,阴性对照组和第3转染组的细胞活力分别为(1.01±0.02)%,(0.72±0.05)%;这2组的细胞凋亡率分别为(34.66±0.94)%,(56.71±3.03)%,组间比较差异均有统计学意义(均P<0.05)。miR-876-3p可靶向结合PPIA。以1μg·mL^-1顺铂处理MKN28细胞,阴性对照组、第1转染组和第3转染组的细胞活性分别为(1.01±0.03)%,(0.58±0.04)%和(0.97±0.02)%;这3组的细胞凋亡率分别为(39.96±2.01)%,(71.12±2.98)%和(45.14±2.47)%。以5μg·mL^-1顺铂处理MKN28/DDP细胞,这3组的细胞活性分别为(1.01±0.04)%,(0.63±0.06)%和(0.94±0.05)%;这3组的细胞凋亡率分别为(31.95±1.31)%,(62.88±5.63)%和(36.84±2.11)%。上述指标:第1转染组与阴性对照组比较,差异均�Objective To explore the effect of microRNA-876-3 p(miR-876-3 p)on proliferation and cisplatin resistance of gastric cancer cells through regulating peptidyl-prolyl cis/trans isomerase A(PPIA).Methods MKN28 cells were cultured in vitro,and low concentration gradient method was used to established cisplatin resistance MKN28 cells(MKN28/DDP).Both MKN28 and MKN28/DDP cells were divided into 4 groups:Negative control group(NC),transfection-1 group(si-PPIA),transfection-2 group(miR-876-3 p mimics)and transfection-3 group(si-PPIA+miR-876-3 p inhibitor).MKN28 cells were treated with 1μg·mL^-1 cisplatin,and MKN28/DDP cells were treated with 5μg·mL^-1 cisplatin.The relative expression of miR-876-3 p was measured by quantitative real time-PCR.The relative expression of PPIA protein was measured by Western blotting.The viability of cells was measured by CCK-8 assay.The apoptosis of cells was measured by flow cytometry assay.Results The relative expression of miR-876-3 p in normal gastric epithelial cells GES-1,gastric cancer cells MKN28 and MKN28/DDP were 1.01±0.21,0.41±0.15 and 0.32±0.11;comparison between MKN28 and GES-1,the difference was significant(P<0.01),and comparison between MKN28/DDP and MKN28,the difference was significant(P<0.05).In over-expression of miR-876-3 p experiments,MKN28 cells were treated with 1μg·mL^-1 cisplatin,the cell viability in transfection-1 group and transfection-3 group were(1.00±0.04)%and(0.65±0.03)%;the apoptosis rate in the two groups were(41.87±1.65)%and(68.98±4.12)%,comparison between transfection-1 group and transfection-3 group,the difference of the factors were significant(all P<0.05);and MKN28/DDP cells were treated with 5μg·mL^-1 cisplatin,the cell viability in two groups were(1.01±0.02)%and(0.72±0.05)%;the apoptosis rate in the two groups were(34.66±0.94)%and(56.71±3.03)%,comparison between transfection-1 group and transfection-3 group,the difference of the factors were significantly(all P<0.05).PPIA is one of the target genes of miR-876-3 p.MKN28 cells were tre

关 键 词：微小RNA-876-3p 肽基脯氨酰顺反异构酶A 胃癌 增殖 顺铂耐药 

分 类 号：R97[医药卫生—药品]