Sox2下游调控蛋白的筛选及其对大肠癌细胞增殖、迁移的影响

摘要

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.%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。