摘要
目的通过siRNA干扰沉默结肠癌转移相关基因1（metastasis．associatedincoloncanceY1，MACCI），观察其对食管癌细胞株TE-1增殖、迁移及侵袭能力的影响。方法通过化学合成的特异性siRNA与阳离子脂质体Lipofectamine TM 2000形成复合体，转染食管癌细胞株TE-1，应用qRT-PCR、Westernblot检测转染后MACCl基因和蛋白表达水平；MTT法检测细胞生长增殖水平；划痕实验、Transwell实验检测细胞株干扰前后迁移侵袭能力的改变。结果siRNA．MACCl转染TE一1细胞后，与空白组比较，干扰组MACCl基因和蛋白的表达水平明显降低，分别降低71％和87％；MTT实验观察1—7d，干扰组较空白组、阴性对照组增殖速度明显降低（P〈0．05），第4天后效果更明显；划痕实验结果显示干扰组迁移划痕处的细胞（10．6±2．2）明显少于空白组（68．2±2．5），差异具有统计学意义（P〈0．05），各对照组间无统计学差异（P〉0．05）；Transwell实验结果显示干扰组穿过膜的TE-1细胞数目（4．2±1．9）较空白组（58．4±7．4）明显减少（P〈0．05），各对照组间无统计学差异（P〉0．05），表明干扰后TE-1细胞的迁移侵袭能力显著降低。结论siRNA．MACCl干扰导致TE-1细胞MACCl基因及蛋白表达水平明显下调；MACCl基因下调后，食管癌TE-1细胞的增殖、迁移、侵袭能力均明显降低，说明MACCl基因具有促进食管癌细胞增殖、迁移、侵袭的能力。
Objective To determine the effects of siRNA interfering metastasis-associated in colon cancer 1 （MACC1） on the extracorporeal proliferation, migration and invasion of the esophageal squamous cell carcinoma （ESCC） cell line TE-1. Methods The special MACCI-siRNA sequence was chemically synthesize, and then transfected into the TE-1 cells with lipofectamineTM 2000. RT-quantity PCR and Western blotting were used to detect the mRNA and protein level of MACC1 in untreated TE-1 cells, lipo2000-treated cells （mock）, siRNA-nc transfected cells, and siRNA-MACC1 transfected cells. MTI＂ assay was used to detect the proliferation of TE-1 ceils. Scratching test and Transwell chamber assay were evaluated the migration of the cells. Results Compared with the untreated cell, mock, and siRNA-nc cells, the expression of MACC1 was obviously decreased by 71% at mRNA and 87% at protein levels. In 7 dafter transfection, MTT assay indicated that the proliferation of the transfected TE-1 ceils was significantly inhibited compared with the other 3 types of ceils （ P 〈 0.05 ）, especially from the fourth day. Scratching test showed that there were obviously less migrated cells from the siRNA-MACC1 group than those from the control （ 10.6 ± 2.2 vs 68.2 ± 2.5, P 〈 0.05 ）. Transwell assay also showed the invasion ability of TE-1 ceils transfected with siRNA-Maccl was lesser than the control cells （4.2 ± 1.92 vs 58.4 ± 7.44, P 〈 O. 05 ）. But no significant difference was seen among the untreated cell, mock, and siRNA-nc cells. Conclusion siRNA interference of MACC1 obviously down-regulates the molecule at mRNA and protein levels in TE-1 cells, and results in significant inhibition in the proliferation, migration and invasion of the cells. Therefore, MACC1 might be involved in the growth, migration and invasion of ESCC.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2014年第5期442-445,共4页
Journal of Third Military Medical University
基金
四川省教育厅重点项目(11ZA148)~~
关键词
食管鳞癌
结肠癌转移相关基因1
RNA
小分子干扰
细胞增殖
细胞迁移
肿瘤侵袭
esophageal squamous cell carcinoma
metastasis-associated in colon cancer 1
RNA, smallinterfering
cell proliferation
cell movement
neoplasm invasiveness
