蒲公英提取物调控DPC4对大肠癌细胞凋亡、迁移、侵袭的影响及其机制研究

Study on Effect of Herba Taraxaci Extract Regulating DPC4 on Apoptosis,Migration and Invasion of Colorectal Cancer Cells and Its Mechanism

目的:探讨蒲公英提取物通过调控胰腺癌缺失位点4(DPC4)表达对大肠癌细胞凋亡、迁移和侵袭的影响和机制。方法:用不同终浓度(1.5、3.0、7.5、15.0 mg/mL)的蒲公英提取物处理HT-29细胞48 h后,流式细胞术检测细胞凋亡,Transwell法检测细胞的迁移和侵袭能力,Western blot检测DPC4蛋白的表达。将pcDNA3.1-DPC4转染HT-29细胞,观察过表达DPC4对HT-29细胞凋亡、迁移和侵袭的影响。将si-DPC4转染HT-29细胞,给予蒲公英提取物处理后,观察抑制DPC4表达对HT-29细胞凋亡、迁移和侵袭的影响。结果:蒲公英提取物在体外可诱导HT-29细胞凋亡,抑制HT-29细胞的迁移和侵袭,并呈明显的浓度依赖性。15.0 mg/mL的蒲公英提取物可促进DPC4蛋白的表达。过表达DPC4可显著促进HT-29细胞凋亡,抑制HT-29细胞的迁移和侵袭。抑制DPC4表达能逆转蒲公英提取物对HT-29细胞凋亡、迁移和侵袭的影响。结论:蒲公英提取物通过上调DPC4表达进而促进大肠癌细胞凋亡,抑制细胞的迁移和侵袭能力。

objective:To investigate the effect and mechanism of Herba Taraxaci extract on apoptosis,migration and invasion of colorectal cancer cells by regulating the expression of deleted in pancreatic carcinoma locus 4(DPC4).Methods:After treating HT-29 cells with Herba Taraxaci extract of different final concentrations including 1.5,3.0,7.5 and 15.0 mg/mL for 48 hours,cell apoptosis was detected by flow cytometry,the ability of migration and invasion of cells was detected by Transwell method,and expression of DPC4 protein was detected by Western blot.PcDNA3.1-DPC4 was transfected into HT-29 cells,and the effect of over-expression of DPC4 on apoptosis,migration and invasion of HT-29 cells was observed.Si-DPC4 was transfected into HT-29 cells,and the effect of the inhibition of DPC4 expression on apoptosis,migration and invasion of HT-29 cells was observed after being treated with Herba Taraxaci extract.Results:Herba Taraxaci extract in vitro could induce the apoptosis of HT-29 cells and inhibit the migration and invasion of HT-29 cells in an obvious concentrationdependent manner.Fifteen mg/mL of Herba Taraxaci extract can promote the expression of DPC4 protein.Over-expression of DPC4 could significantly promote the apoptosis of HT-29 cells and inhibit the migration and invasion of HT-29 cells.Inhibition of DPC4 expression could reverse the effect of Herba Taraxaci extract on apoptosis,migration and invasion of HT-29 cells.Conclusion:By up-regulating the expression of DPC4,Herba Taraxaci extract can promote the apoptosis of colorectal cancer cells and inhibit the ability of migration and invasion of cells.