腺病毒介导miRNA-29a过表达抑制人胃癌细胞的增殖  

Adenovirus mediated over-expression of miRNA-29a inhibits proliferation of human gastric cancer cells

作　　者：刘志鹏[1] 王宗华[2] 卢斌[3] 胡春燕[3] 李学成[1] 邹利全[1] 张方征[1] 陈陵[1] 

机构地区：[1]解放军第324医院消化内科,重庆400020 [2]解放军第三军医大学护理学院基础护理学教研室,重庆400038 [3]解放军第三军医大学新桥医院肿瘤生物治疗中心,重庆400037

出　　处：《中国肿瘤生物治疗杂志》2013年第3期312-316,共5页Chinese Journal of Cancer Biotherapy

基　　金：重庆市自然科学基金资助项目(No.CSTC;2010BB5158;No.CSTC;2011AC5024;No.CSTC;2011jjA0878;No.CSTC;2011jjA0882)~~

摘　　要：目的:利用重组人5型复制缺陷型腺病毒载体介导miRNA-29a(miR-29a)过表达,观察miR-29a对人胃癌细胞系SGC-7901及AGS的增殖抑制作用。方法:构建含pre-miR-29a的重组腺病毒Ad-miR29a及含LacZ基因的对照腺病毒Ad-LacZ,分别感染人胃癌SGC-7901及AGS细胞,采用real-time PCR法检测SGC-7901及AGS细胞中miR-29a的表达,CCK-8法检测miR-29a对SGC-7901及AGS细胞增殖的抑制作用,流式细胞术检测SGC-7901及AGS细胞的细胞周期,Transwell法检测miR-29a对SGC-7901及AGS细胞迁移能力的影响。结果:与Ad-LacZ对照组相比,Ad-miR29a组感染后胃癌SGC-7901及AGS细胞中miR-29a的表达均显著增加[(17.35±0.71)vs(1.12±0.09),(26.50±1.09)vs(0.95±0.04);均P<0.01];且胃癌SGC-7901及AGS细胞增殖明显受到抑制,感染6 d后D值均显著降低[(0.54±0.03)vs(0.77±0.04),(0.70±0.03)vs(0.88±0.04);均P<0.01)],而Ad-LacZ感染组与未感染组相比则无明显变化(P>0.05);与Ad-LacZ对照组相比,Ad-miR29a组SGC-7901及AGS细胞中G0/G1期细胞比例均显著增加[(63.10±4.91)%vs(47.60±5.31)%,(69.80±3.15)%vs(54.60±4.22)%;均P<0.05],但胃癌SGC-7901及AGS细胞的迁移能力并没有明显差异。结论:miR-29a过表达可有效抑制胃癌SGC-7901及AGS细胞的增殖,miR-29a有望成为治疗胃癌的新靶标。Objective：To explore the inhibitory effect of miRNA-29a（miR-29a） on the proliferation of human gastric cancer cell lines SGC-7901 and AGS through over-expression of miR-29a mediated by the recombinant replication-deficient human adenovirus type 5 vector.Methods： The recombinant adenovirus Ad-miR-29a containing pre-miR-29a or control adenovirus Ad-LacZ containing LacZ gene was constructed and infected into human gastric cancer SGC-7901 and AGS cells,respectively.The expressions of miR-29a in SGC-7901 and AGS cells were detected by real-time PCR.CCK-8 assay was employed to examine the inhibitory effect of miR-29a on the proliferation of human gastric cancer cell lines.The flow cytometry assay was used to analyze cell cycle of SGC-7901 and AGS cells.The migration ablilities of SGC-7901 and AGS cells were assessed by Transwell assay.Results： Compared with the Ad-LacZ group,the Ad-miR29a group expressed higher level of miR-29a in both SGC-7901 and AGS cells（[17.35±0.71] vs [1.12±0.09],[26.50±1.09] vs [0.95±0.04],P0.01）.The proliferation of SGC-7901 and AGS was significantly inhibited in the Ad-miR29a group as compared with that in the Ad-LacZ group.Six days after Ad-miR29a infection,the D value was significantly decreased（[0.54± 0.03] vs [0.77 ± 0.04],[0.70 ± 0.03] vs [0.88 ± 0.04],P0.01）.The proliferation of cells in the Ad-LacZ group showed no significant changes as compared with that in the uninfected group（P0.05）.Meanwhile,The percentage of SGC-7901 or AGS cells arrested in G0/G1 period in the Ad-miR29a group was significantly higher than that in the Ad-LacZ group（[63.10±4.91]% vs [47.60±5.31]%,[69.80±3.15]% vs [54.60±4.22]%,P0.05].However,no significant changes were found in the migration ability of gastric cancer SGC-7901 and AGS cells.Conclusion： miR-29a can effectively suppress the proliferation of human gastric cancer cells,which makes a promising new therapeutic target for gastric cancer.

关 键 词：miRNA-29a 腺病毒 胃癌 增殖 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤] R730.54[医药卫生—临床医学]