天津医药

• 细胞与分子生物学 • 上一篇    下一篇

siRNA抑制Mcl-1基因表达对胃癌细胞MGC-803生物学行为的影响

刘金禄,王震,陈俊强,崔希刚,马鹏飞,黎伯培   

  1. 广西医科大学第一附属医院
  • 收稿日期:2013-07-15 修回日期:2013-12-23 出版日期:2014-04-15 发布日期:2014-04-15
  • 通讯作者: 陈俊强

The Effect of Silencing Mcl-1by siRNA on Biological Behavior of Gastric Cancer MGC-803Cell

  • Received:2013-07-15 Revised:2013-12-23 Published:2014-04-15 Online:2014-04-15

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摘要： 目的 探讨siRNA抑制Mcl-1基因表达后对胃癌细胞MGC-803生物学行为的影响。方法 合成针对Mcl-1的靶向siRNA（Mcl-1 siRNA组），以空白细胞（空白对照组）和分别转染脂质体试剂（脂质体对照组）及阴性对照siRNA（阴性对照组）作为对照组。噻唑蓝（MTT）检测Mcl-1 siRNA对MGC-803细胞增殖能力的影响；流式细胞术观察各组细胞凋亡及周期变化情况；转染48 h后应用Transwell小室分析细胞侵袭、迁移能力的变化。结果 Mcl-1 siRNA组转染后24 h、48 h、72 h的A值低于3个对照组（P＜0.05），其细胞增殖抑制率分别为8.9%、21.6%和18.8%。转染48 h后，Mcl-1 siRNA组细胞凋亡率高于空白对照组、脂质体对照组和阴性对照组（%：19.61±1.66 vs 3.69±0.37 vs 3.54±0.47 vs 3.68±0.55，F=12.230，P＜0.05）。Mcl-1 siRNA组G0/G1[（41.03±1.86）%]、G2/M期比例[（1.80±0.46）%]均低于3个对照组，S期比例[（57.17±1.72）%]高于3个对照组（均P＜0.05）。Mcl-1 siRNA组侵袭、迁移实验中的穿膜细胞数（分别是42.00±4.00、76.33±3.51）均低于空白对照组（分别是79.33±3.51、108.00±3.61）、脂质体对照组（分别是74.67±2.52、110.67±4.04）和阴性对照组（分别是77.33±3.06、109.33±4.51）。以上指标在3个对照组间差异均无统计学意义。结论 抑制Mcl-1表达可有效降低胃癌细胞的增殖、侵袭及迁移能力，并促进肿瘤细胞凋亡。

关键词: 胃肿瘤, RNA, 小分子干扰, 细胞增殖, 细胞凋亡, 细胞侵袭, 细胞迁移, siRNA

Abstract: Objective To investigate the effect of inhibiting Mcl-1 gene expression on the biological behavior of gastric cancer cell MGC-803 by using a small interference RNA(siRNA)strategy. Methods Synthesized siRNA targeting Mcl-1(Mcl-1siRNA group) was transfected into MGC-803 cells,additionally,MGC-803 cells transfected with negative siRNA(Mcl-1siRNA-NC group),MGC-803 cells transfected with Lipofectamine 2000（liposomes control group）and vacant MGC-803 cells（blank control group） were used as controls.MTT was adapted to investigate the proliferation of MGC-803 cells after transfection of Mcl-1 siRNA. After 48h transfection of Mcl-1 siRNA, the flow cytometry(FCM) was used to examine the apoptosis cells in the four groups. Cell cycle was detected by FCM in different groups. Polycarbonate membrane transwell chamber was used for detecting the abilities of invasion and migration of the cell line. Results The absorption value of MGC-803 cells decreased greatly after transfected with Mcl-1siRNA for 24、48 and 72 h compared to those in control groups(P<0.05); after transfected 48h, apoptosis rate in Mcl-1siRNA group was higher than in the blank control group, liposomes control group and Mcl-1siRNA-NC group(%:19.61±1.66 vs 3.69±0.37 vs 3.54±0.47 vs 3.68±0.55,F=12.230,P＜0.05),G0/G1 [(41.03±1.86)%] and G2 / M phase ratio [(1.80± 0.46)%] in Mcl-1 siRNA group were lower than in the three control groups, S phase ratio [(57.17±1.72)%] was higher than in three control groups(P<0.05). Transmembrane cell number of Mcl-1 siRNA group in polycarbonate membrane transwell chamber(42.00±4.00,76.33±3.51 respectively)were less than of the blank control group(79.33±3.51,108.00±3.61 respectively),liposome control group(74.67±2.52,110.67±4.04 respectively)and negative control group (77.33±3.06,109.33±4.51 respectively). However,there was no significant difference in above index among the control groups. Conclusion Inhibition the expression of Mcl-1 can effectively supress the growth, invasion and migration, and can promote apoptosis in gastric cancer cells.

Key words: Stomach Neoplasms, RNA, Small Interfering, Cell Proliferation, Cell Apoptosis, Cell invasion, Cell Migration, siRNA