摘要
目的探讨mi R-711对胃癌MGC-803细胞侵袭转移及上皮间质转化(epithelial mesenchymal transition,EMT)的影响。方法通过脂质体瞬时转染法将mi R-711过表达(mi R-711 mimics)、mi R-711抑制表达(mi R-711 inhibitors)、mi RNA空质粒转染人胃癌细胞株MGC-803。以mi RNA空质粒转染组作为阴性对照组(NC组),以空白转染组作为空白对照组(K组)。转染48 h后在荧光显微镜下观察转染效率,然后采用Transwell侵袭小室实验检测细胞的侵袭能力,细胞划痕实验检测细胞的迁移能力,Western blot实验检测上皮蛋白和间质蛋白表达量的变化。结果 Transwell侵袭实验结果显示:与阴性对照组穿膜细胞数(120.00±4.58)及空白对照组穿膜细胞数(119.67±5.13)分别比较,mi R-711 mimics组穿膜细胞数(70.67±5.51)明显下降(P均<0.05);划痕实验显示:划痕48 h,与阴性对照组细胞愈合率(32.52±1.73)%及空白对照组细胞愈合率(32.15±1.55)%分别比较,mi R-711mimics组细胞愈合率(7.08±0.73)%明显下降(P均<0.05);Western blot实验显示:mi R-711 mimics组与阴性对照及空白对照组分别比较,上皮标志物(E-cadherin蛋白)相对蛋白表达量增多,差异有统计学意义(P均<0.05);间质标志物(vimentin蛋白)相对蛋白表达量水平减少,差异有统计学意义(P均<0.05)。结论过表达mi R-711可抑制胃癌MGC-803细胞侵袭迁移及上皮间质转化的发生。
Objective To investigate the effects of mi R-711 on the invasion, metastasis and epithelial mesenchymal transition（EMT） of human gastric cancer cells MGC-803. Methods mi R-711 mimics, mi R-711 inhibitors and mi RNA empty plasmids were transfected into human gastric cancer cell strain MGC-803 by liposomes transient transfection method. mi RNA empty plasmids transfection group was taken as the negative control group（NC group）, and the blank transfection group taken as blank control group（K group）. After 48 h, the effectiveness of transfection was observed under fl uorescence microscope. Cell scratch experiment and Transwell invasion experiment were used to detect the migration and invasion of transfected MGC-803 cells. Western blot was used to observe the expression of epithelial protein and mesenchymal protein of transfected MGC-803 cells. Results As revealed by Transwell invasion test： the number of invaded cells in NC and K groups were（120.00±4.58） and（119.67±5.13）, while that was signifi cantly lower in the mi R-711 mimics transfection group（70.67±5.51）（both P〈0.05）; As revealed by the cell scratch test： at 48 h after scratch, the healing rate of NC and K groups were（32.52±1.73）% and（32.15±1.55）%, while that was signifi cantly lower of the mi R-711 mimics transfection group（7.08±0.73）%（both P〈0.05）; As revealed by Western blot test： compared with NC and K groups, the epithelial markers（E-cadherin） relative protein expression was increased in the mi R-711 mimics transfection group（both P〈0.05）; while the mesenchymal markers（Vimentin） expression was decreased（both P〈0.05）. Conclusion mi R-711 overexpression could inhibit the invasion, metastasis and EMT of human gastric cancer cells MGC-803.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2016年第3期201-206,共6页
Cancer Research on Prevention and Treatment
基金
湖南省教育厅科研项目(13C822)
