南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2014年
11期
1594-1600
,共7页
周敏%陆滟霞%袁理%郑林%刘燕%洪敏%张超%李学农
週敏%陸滟霞%袁理%鄭林%劉燕%洪敏%張超%李學農
주민%륙염하%원리%정림%류연%홍민%장초%리학농
大肠癌细胞%质谱分析%Sox2%调控%S3a%ENO1%增殖%迁移
大腸癌細胞%質譜分析%Sox2%調控%S3a%ENO1%增殖%遷移
대장암세포%질보분석%Sox2%조공%S3a%ENO1%증식%천이
colonic cancer cells%mass spectrometry%Sox2%regulation%S3a%ENO1%proliferation%migration
目的：分析大肠癌细胞转录因子Sox2对大肠癌细胞增殖及迁移的影响。方法应用SDS-PAGE蛋白电泳，结合考马斯亮蓝及镀胺银染色方法，筛选差异蛋白。质谱分析筛选Sox2下游调控蛋白，应用定量PCR（QPCR）及Western blotting验证和鉴定下游调控蛋白。Cell Counting Kit-8（CCK8）观察Sox2对大肠癌细胞增殖的影响，Transwell实验观察Sox2对迁移能力的影响。结果应用蛋白组学技术成功筛选出Sox2下调蛋白S3a、上调蛋白ENO1、Gama-actin。QPCR和Western blotting显示，Sox2过表达后，大肠癌细胞S3a表达减少（P<0.005），ENO1表达增加（P<0.05），Gama-actin表达无明显差异，细胞增殖及迁移能力提高（P<0.05）；Sox2干扰后，大肠癌细胞S3a表达增加（P<0.05），ENO1表达减少（P<0.001），Gama-actin表达无明显差异，细胞增殖及迁移能力下降（P<0.05）。结论 Sox2负向调控S3a的表达，正向调控ENO1的表达，促进大肠癌细胞增殖及迁移。
目的：分析大腸癌細胞轉錄因子Sox2對大腸癌細胞增殖及遷移的影響。方法應用SDS-PAGE蛋白電泳，結閤攷馬斯亮藍及鍍胺銀染色方法，篩選差異蛋白。質譜分析篩選Sox2下遊調控蛋白，應用定量PCR（QPCR）及Western blotting驗證和鑒定下遊調控蛋白。Cell Counting Kit-8（CCK8）觀察Sox2對大腸癌細胞增殖的影響，Transwell實驗觀察Sox2對遷移能力的影響。結果應用蛋白組學技術成功篩選齣Sox2下調蛋白S3a、上調蛋白ENO1、Gama-actin。QPCR和Western blotting顯示，Sox2過錶達後，大腸癌細胞S3a錶達減少（P<0.005），ENO1錶達增加（P<0.05），Gama-actin錶達無明顯差異，細胞增殖及遷移能力提高（P<0.05）；Sox2榦擾後，大腸癌細胞S3a錶達增加（P<0.05），ENO1錶達減少（P<0.001），Gama-actin錶達無明顯差異，細胞增殖及遷移能力下降（P<0.05）。結論 Sox2負嚮調控S3a的錶達，正嚮調控ENO1的錶達，促進大腸癌細胞增殖及遷移。
목적：분석대장암세포전록인자Sox2대대장암세포증식급천이적영향。방법응용SDS-PAGE단백전영，결합고마사량람급도알은염색방법，사선차이단백。질보분석사선Sox2하유조공단백，응용정량PCR（QPCR）급Western blotting험증화감정하유조공단백。Cell Counting Kit-8（CCK8）관찰Sox2대대장암세포증식적영향，Transwell실험관찰Sox2대천이능력적영향。결과응용단백조학기술성공사선출Sox2하조단백S3a、상조단백ENO1、Gama-actin。QPCR화Western blotting현시，Sox2과표체후，대장암세포S3a표체감소（P<0.005），ENO1표체증가（P<0.05），Gama-actin표체무명현차이，세포증식급천이능력제고（P<0.05）；Sox2간우후，대장암세포S3a표체증가（P<0.05），ENO1표체감소（P<0.001），Gama-actin표체무명현차이，세포증식급천이능력하강（P<0.05）。결론 Sox2부향조공S3a적표체，정향조공ENO1적표체，촉진대장암세포증식급천이。
Objective To screen the down-stream proteins of transcription factor Sox2 and explore the role of Sox2 in the proliferation and migration of colonic cancer cells in vitro. Methods The cellular proteins were separated by SDS-PAGE electrophoresis and stained with Coomassie blue and amine plated silver. The differentially expressed proteins was identified by mass spectrometry and verified by QPCR and Western blotting. A cell counting kit-8 (CCK8) assay was performed to evaluate the cell proliferation, and the cell migration was assessed using Transwell assay. Results S3a was identified by proteomics technology as a Sox2-downregulated protein while ENO1 and gama-actin the up-regulated proteins. QPCR and Western blotting analyses showed that overexpression of Sox2 significantly decreased the expression of S3a (P<0.005) and increased the expression of ENO1(P<0.05), but had no significant effect on gama-actin expression. Sox2 overexpression obviously promoted cell proliferation and migration (P<0.05), while inhibition of Sox2 produced contrary effects (P<0.05). Conclusion Sox2 negatively regulates S3a expression and positively regulates ENO1 expression to promot the proliferation and migration of colonic cancer cells.
