摘要
目的探讨抑制自噬基因Beclin1表达对舌鳞状细胞癌(以下简称舌鳞癌)细胞增殖及侵袭转移的影响,了解舌鳞癌侵袭转移的调控机制。方法利用RNAi技术,针对人cDNA序列设计Beclin1干扰序列,用脂质体Lipo2000包裹后转染至舌鳞癌UM2细胞。实验设空白对照组、脂质体2000(Lipo2000)组、阴性siRNA组和Beclin1siRNA组,通过蛋白印迹法检测Beclin1、LC3的表达变化;CCK-8法检测细胞增殖能力变化;Transwell小室检测细胞迁移和侵袭能力改变;SPSS13.0统计软件进行数据分析。结果转染Beclin1siRNA对UM2细胞Beclin1有显著敲减作用(P<0.05),自噬标记蛋白LC3-Ⅱ的表达明显降低(P<0.05),细胞增殖较对照组增快(P<0.05),细胞迁移和侵袭转移能力明显增强。结论沉默Beclin1表达可下调舌鳞癌细胞自噬水平,并促进舌鳞癌细胞增殖和侵袭能力。
Objective To investigate the effect of Beclinl siRNA on proliferation, invasion and migration and their related mechanisms in tongue squamous cell carcinoma （TSCC） cell line. Methods Used the technology of RNA interference （RNAi） to cut down Beclinl expression, targeting the coding sequence of human Beelinl gene, and then transfected into UM2 cells by Lipofectmine 2000. UM2 cells were cultured and divided into 4 groups： blank control group, Lipo2000 control group, nonspecific siRNA group and Beelinl siRNA group. Then, Western blot were performed to examine the expression of Beclinl and LC3. CCK-8 assay were used to evaluate cell proliferation. Transwell assay was carried out to assess cell migration and invasion. Results The Beelinl protein expression was significantly decreased after Beclinl siRNA transfection （P 〈 0.05）. Meanwhile, The expression of LC3-II protein was significantly lower than the control group 72 h after transfection （P 〈 0.05）. Compared to the control group, cell proliferation was significantly increased （P 〈 0.05）. Invasion and migration ability were also enhanced in UM2 cell transfected with Beclinl siRNA （P〈 0.05）. Conclusion Down- regulation of Beelinl can downregulate autophagical activity and promote the proliferation and invasiveness of TSCC cell line.
出处
《中华口腔医学研究杂志（电子版）》
CAS
2013年第1期9-12,共4页
Chinese Journal of Stomatological Research(Electronic Edition)
基金
广东省自然科学基金(10151008901000025)
广东省科技计划(2009B050700024
2009B060700096)
广州市科技计划项目对外科技合作专项(2012J5100008)
