天津医药

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上调miR-200a、miR-141表达对胃腺癌细胞生长的体外研究

申发娟1,苏娟2,张庆瑜3,康春生4   

  1. 1. 天津医科大学总医院
    2. 天津医科大学总医院耳鼻喉科
    3. 天津医科大学总医院消化内科
    4. 天津医科大学总医院神经病研究所
  • 收稿日期:2012-07-24 修回日期:2013-02-03 出版日期:2013-06-15 发布日期:2013-06-15
  • 通讯作者: 张庆瑜

Effects of up-regulated miR-200a and miR-141 on gastric adenocarcinoma cell growth in vitro

SHEN Fajuan 1,SU Juan 2,ZHANG Qing yu1,KANG Chun sheng3   

  1. 1. Department of Gastroenterology, General Hospital of Tianjin Medical University
    2. Department of Otolaryngology
    3. Neurology Institute
  • Received:2012-07-24 Revised:2013-02-03 Published:2013-06-15 Online:2013-06-15
  • Contact: ZHANG Qing yu

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摘要： 目的  探究上调microRNA（miR）-200a和miR-141表达对胃腺癌细胞系SGC-7901细胞侵袭迁移的影
响。方法  脂质体介导miR-200a（miR-200a组）、miR-141（miR-141组）、miR-200a+miR-141（miR-200a+miR-141组）拟似物转染SGC-7901细胞，同时设未转染组(对照组)、无义序列转染组。应用qRT-PCR检测转染后各组miR的表达，Transwell小室法和划痕实验检测转染后细胞侵袭、迁移能力的变化，Western blot检测转染后E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（N-cadherin）、基质金属蛋白酶-9（MMP-9）的表达，并进行比较分析。结果   与对照组和无义序列转染组相比，miR-200a、miR-141 和miR-200a+miR-141 组miR 表达较高，细胞穿膜细胞数较少（均P <0.05），划痕法显示SGC-7901细胞阻滞效果更明显，E-cadherin表达增加，N-cadherin、MMP-9表达减少，且以上变化在miR-200a+miR-141组更显著。结论   上调miR-200a、miR-141表达可有效抑制SGC-7901细胞的侵袭迁移。

关键词: 微RNAs, 胃肿瘤, 腺癌, 肿瘤细胞, 培养的, 肿瘤侵润, 肿瘤转移, 钙黏着糖蛋白类

Abstract: Objective    To investigate the effects of up-regulated expression of microRNA(miR)-200a and miR-141 on the invasion and migration capability of SGC7901 human gastric adenocarcinoma cells. Methods   Synthetic miR-200a and miR-141 mimics was transfected into SGC7901 cell line by lipofectamine 2000; control group(without transfection), group B(perfomling transfection with nonsense sequence)and group E(performing transfecfion with both miR-200a and miR-141 mimics)were established．Realtime quantitative PCR was conducted to detect the expressions of miR-200a and miR-141 in these groups after transfection．The migration，invasion capability of cells were evaluated by using transwell chamber model and wound healing assay in vitro, while the expression of E-cadherin, N-cadherin, MMP-9 were determined by Western blot. Results    As compared with those in control group and group B，the expressions of miR-200a , miR-141and E-cadherin increased significantly(P<0.05). The mobility，invasion，migration capability and the expression of MMP-9, N-cadherin proteins of the cells transfected with miRNA mimics were much lower than the other two groups(P<0.05), while the differences between the other two groups were not significant. Conclusion    The up-regulated expression of miR-200a and miR-141 could significantly inhibit the invasion and migration ability of SGC7901 cells.

Key words: microRNAs, stomach neoplasms, tumor cells, cultured, neoplasm invasiveness, cadherins, matrix metalloproteinase-9, 钙黏着糖蛋白类