丙戊酸对食管癌细胞增殖凋亡的影响及其抗肿瘤活性机制的研究

Effect of Valproic Acid on proliferation and apoptosis of esophageal cancer cells and study on its antitumor activity mechanism

目的探索丙戊酸(VPA)对食管癌细胞增殖凋亡的影响以及Bcl-2蛋白、Caspase蛋白表达及信号转导通路的分子机制。方法将食管癌ECa-109细胞株培养于PRMI1640培养液,按照溶剂对照和溶度梯度分组:空白对照及VPA 0.25 mmol/L、0.5 mmol/L、1.0 mmol/L、2.0 mmol/L、4.0 mmol/L。根据不同的药物浓度处理细胞24、48、72 h后,观察食管癌ECa-109细胞的变化,运用不同的检测方法检测食管癌ECa-109细胞的细胞活力、细胞凋亡及周期的变化;Western Blot检测CyclinD1、p21、Survivin、Bcl-2和Caspase蛋白及PI3K/Akt和MAPK信号转导通路的变化。结果VPA可有效影响ECa-109细胞的活力、增殖和分化,能够通过诱导细胞周期阻滞使细胞产生凋亡。Western Blot实验结果显示,VPA可以影响ECa-109细胞中Bcl-2、Survivin以及Caspase蛋白的表达;VPA可明显降低ECa-109细胞中CyclinD1蛋白表达水平,同时增加p21蛋白的表达,使其能够影响信号通路PI3K/Akt和MAPK途径中AKT、MAPK等关键蛋白的磷酸化。结论VPA能够影响食管癌细胞的增殖凋亡,并具有浓度依赖性;能够通过影响多种蛋白激酶途径中的信号通路、生存蛋白的表达以及PI3K/Akt和MAPK信号通路的表达而影响食管癌细胞的生长。

Objective To explore the effect of Valproic Acid(VPA)on proliferation and apoptosis of esophageal cancer cells,and expression of Bcl-2,Caspase protein,as well as molecular mechanisms of signal transduction pathways.Methods ECa-109 cell line of esophageal cancer was cultured with PRMI1640 medium,and grouped according to solvent control and solubility gradient:blank control and VPA 0.25 mmol/L,0.5 mmol/L,1.0 mmol/L,2.0 mmol/L,4.0 mmol/L.After treating cells for 24,48 and 72 hours according to different drug concentrations,the changes of esophageal cancer ECa-109 cells were observed,and different detection methods were used to detect the changes of cell viability,apoptosis and cycle of esophageal cancer ECa-109 cells.Western Blot was used to detect CyclinD1,p21,Survivin,Bcl-2,Caspase protein and PI3K/Akt and MAPK signal transduction pathway changes.Results VPA showed different extent changes on vitality,proliferation,differentiation of ECa-109 cells,could induce cell apoptosis by inducing cell cycle arrest.Western Blot experiment results showed that VPA could affect the expression of Bcl-2,Survivin and Caspase proteins in ECa-109 cells.VPA could significantly reduce the expression level of CyclinD1 protein in ECa-109 cells and increase the expression of p21 protein,which influenced the phosphorylation of key proteins such as AKT and MAPK in the signaling pathway PI3K/Akt and MAPK pathway.Conclusion VPA has an effect on the proliferation and apoptosis of esophageal cancer cells and it is concentration-dependent,which can affect the growth of esophageal cancer cells through the signaling pathways of multiple protein kinase pathways,the expression of survival proteins,the expression of PI3K/Akt and MAPK signaling pathways.