miRNA-214对胃癌细胞BGC823,MKN45的细胞周期和凋亡的影响  

The effect of miRNA-214 on cell cycle and apoptosis of gastric cancer cell lines BGC823 and MKN45

作　　者：熊欣[1] 郑长黎[1] 

机构地区：[1]中南大学湘雅医学院病理学系,湖南长沙410013

出　　处：《中国普通外科杂志》2010年第12期1311-1315,共5页China Journal of General Surgery

摘　　要：目的探讨miRNA-214对胃癌细胞的细胞周期和凋亡的影响,及其对靶基因PTEN蛋白表达的影响。方法荧光定量PCR法检测人低分化胃癌细胞BGC823、MKN45和人正常胃黏膜细胞GES-1中miRNA-214的表达;检测瞬时转染反义核苷酸(miRNA-214抑制剂)后的miRNA-214的表达;流式细胞术分析转染后的细胞生长周期和凋亡率的变化;免疫荧光检测miRNA-214抑制剂对靶基因PTEN表达的影响。结果 miRNA-214在人胃癌细胞BGC823及MKN45中表达上调(P<0.05)。转染miRNA-214抑制剂后能使BGC823中miRNA-214的表达下调(P<0.05),且BGC823和MKN45在转染组的G1期细胞较未转染组明显增高[分别为(60.20±3.38)%vs.(49.33±7.99)%(P<0.05);(69.90±0.28)%vs.(54.85±0.64)%(P<0.05)],S期细胞在转染组较未转染组明显降低[(21.87±3.20)%,(18.25±1.34)%,P<0.05)],但对细胞凋亡率无影响(P>0.1)。免疫荧光显示,转染miRNA-214抑制剂后PTEN呈强表达。结论 (1)人胃癌细胞系BGC823,MKN45中的miRNA-214呈高表达;(2)miRNA-214可使这两株细胞的G1期细胞减少,S期细胞增多,下调PTEN可能是其作用机制之一;(3)miRNA-214对上述两株细胞的凋亡无影响。Objective To identify the effects of miRNA-214 on cell cycle and apoptosis of gastric cancer cells,and its influence on PTEN protein level. Methods miRNA-214 in poorly differentiated human gastric cancer cell line BGC823,MKN45 and normal gastric mucosal cell line GES-1 was detected by real-time PCR. Antisense-miRNA-214 oligonucleotides（miRNA-214 inhibitor） were transfected transiently into gastric cancer cell lines to down-regulate the expression of miRNA-214. The cell cycle and apoptosis rate were studied by flow cytometry. PTEN,the target gene of miRNA-214,was detected by immunofluorescence. Results The miRNA-214 was upregulated in gastric cancer cell BGC823 and MKN45 （P0.05） compared with normal gastric mucosal cell line GES-1. The transfection of miRNA-214 inhibitor downregulated miRNA-214 expression in BGC823 （P0.05）. G1-phase cells were increased in BGC823 and MKN45,[（60.20±3.38）% vs. （49.33±7.99）% （P0.05）;（69.90±0.28） vs. （54.85±0.64）% （P0.05） respectively] compared with the control groups. But the alteration of apoptotic rate was not significant（P0.1）. The immunofluorescence result showed that PTEN was upregulated after transfection in both cell lines. Conclusions （1）miRNA-214 is upregulated in human gastric cancer cell BGC823 and MKN45. （2）miRNA-214 could reduce the G1-phase cells and increase S-phase cells in BGC823 and MKN45,and the downregulation of PTEN may be one of the mechanisms. （3）miRNA-214 has no effect on apoptosis of BGC823 and MKN45.

关 键 词：miRNA 胃癌细胞 细胞周期 凋亡率 PTEN 

分 类 号：R73[医药卫生—肿瘤]