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siRNA干扰Mig-7基因对人胃癌MKN45细胞侵袭和迁移的影响

陈勇华, 高青, 李文丽

陈勇华, 高青, 李文丽. siRNA干扰Mig-7基因对人胃癌MKN45细胞侵袭和迁移的影响[J]. 肿瘤防治研究, 2015, 42(10): 979-983. DOI: 10.3971/j.issn.1000-8578.2015.10.006
引用本文: 陈勇华, 高青, 李文丽. siRNA干扰Mig-7基因对人胃癌MKN45细胞侵袭和迁移的影响[J]. 肿瘤防治研究, 2015, 42(10): 979-983. DOI: 10.3971/j.issn.1000-8578.2015.10.006
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CHEN Yonghua, GAO Qing, LI Wenli. Effect of siRNA Targeting Migration-inducing Protein 7 Gene on Invasion and Migration of Human Gastric Cancer Cells MKN45[J]. Cancer Research on Prevention and Treatment, 2015, 42(10): 979-983. DOI: 10.3971/j.issn.1000-8578.2015.10.006
Citation: CHEN Yonghua, GAO Qing, LI Wenli. Effect of siRNA Targeting Migration-inducing Protein 7 Gene on Invasion and Migration of Human Gastric Cancer Cells MKN45[J]. Cancer Research on Prevention and Treatment, 2015, 42(10): 979-983. DOI: 10.3971/j.issn.1000-8578.2015.10.006
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siRNA干扰Mig-7基因对人胃癌MKN45细胞侵袭和迁移的影响

  • 1. 315202 宁波，宁波市镇海区人民医院消化内科；2. 400016 重庆，重庆医科大学附属第一医院消化内科

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    作者简介:

    陈勇华（1976-），男，本科，副主任医师，主要从事消化内科治疗工作

  • 中图分类号: R735.2

Effect of siRNA Targeting Migration-inducing Protein 7 Gene on Invasion and Migration of Human Gastric Cancer Cells MKN45

  • 1. Department of Gastroenterology, Zhenhai District People's Hospital of Ningbo, Ningbo 315202, China; 2. Department of Gastroenterology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China

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    摘要
  • 摘要: 目的 探讨小干扰RNA（small interfering RNA，siRNA）抑制迁移诱导基因7（Mig-7）基因表达，对人胃癌MKN45细胞侵袭和迁移的影响及可能机制。方法 化学合成靶向Mig-7基因的siRNA，脂质体法转染MKN45细胞，分别用real-time PCR和Western blot法验证其沉默效率；MTT法检测细胞的增殖情况；Transwell小室方法检测细胞侵袭及迁移能力；Western blot法检测基质金属蛋白酶2（MMP-2）和MMP-9蛋白表达。结果 化学合成siRNA转染胃癌MKN45细胞后，Mig-7的mRNA 水平下降（80±2.91）%，蛋白表达水平下降（89.1±2.67）%；沉默Mig-7基因后，各组细胞体外生长速度差异无统计学意义（P>0.05），但侵袭和迁移能力下降，MMP-2蛋白表达水平下降，与阴性转染组和对照组相比差异有统计学意义（P=0.000）。而各组MMP-9蛋白表达水平差异无统计学意义（P>0.05）。结论 下调人胃癌MKN45细胞中Mig-7的表达，可能通过下调MMP-2蛋白表达从而抑制细胞的侵袭和迁移能力。

     

    Abstract: Objective To investigate the effect of small interfering RNA(siRNA) targeting migrationinducing protein 7(Mig-7) gene on the invasion and migration abilities of human gastric cancer cells MKN45, and the underlying mechanism. Methods The chemically synthetic siRNA targeting Mig-7 gene was transfected into MKN45 cells via LipofectamineTM 2000. The gene silencing effect of siRNA targeting Mig-7 was determined by real-time PCR and Western blot at mRNA and protein levels, respectively. Cell proliferation was examined by MTT assay. The cell invasion and migration abilities were determined by Transwell experiment. The protein levels of matrix metalloproteinase-2(MMP-2) and MMP-9 were analyzed by Western blot. Results After transfection of chemically synthetic siRNA into MKN45 cells, the mRNA and protein levels of Mig-7 were significantly reduced by (80±2.91)% and (89.1±2.67)%, respectively. No statistical difference was observed among each group in the growth rates in vitro (P>0.05). Compared with the control groups, the knockdown of Mig-7 in MKN45 cells resulted in significant reduction of cell invasion and migration abilities (P<0.05); down-regulation of Mig-7 in MKN45 cells also reduced the expression of MMP-2 protein (P<0.05). But there was no statistical difference in the expression of MMP-9protein among each group (P>0.05). Conclusion The invasion and migration abilities of gastric cancer cells MKN45 are inhibited by reducing the expression of Mig-7 which might be mediated by downregulating the expression of MMP-2 protein.

     

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出版历程
  • 收稿日期:  2014-10-22
  • 修回日期:  2015-06-10
  • 刊出日期:  2015-10-24

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