启膈散及其拆方抑制原代培养食管癌细胞PDGFR-PLC-γ1酪氨酸的磷酸化

Inhibitory effects of Qigesan and its individual components on tyrosine phosphorylation of PDGFR-PLC-γ1 in primary cultured esophageal carcinoma cells

目的:观察启膈散及其活血和化痰2个拆方对原代培养的食管癌细胞血小板衍生生长因子受体(PDGFR)和磷脂酶C-γ1(PLC-γ1)酪氨酸磷酸化的影响,以探讨启膈散治疗食管癌的作用机制.方法:从外科切除的原发性食管癌组织中采集食管上皮细胞进行原代培养,加入PDGF和启膈散及其活血和化痰2个拆方的水提物,免疫印迹法(Western blotting)测定PDGFR- PLC-γ1蛋白表达和酪氨酸磷酸化水平.结果:用PDGF-BB刺激4 min和启膈散及其拆方处理15 min前后,原代培养食管癌细胞PDGFRβ和PLC-γ1蛋白表达没有变化;但PDGF-BB刺激后PDGFRβ和PLC-γ1蛋白酪氨酸磷酸化明显增强,启膈散及其拆方能不同程度地降低其蛋白磷酸化水平,其中以活血组作用最好,全方组次之.结论:食管癌病变与PDGFR-PLC-γ1蛋白酪氨酸磷酸化增强有关,启膈散及其拆方通过抑制PDGFR-PLC-γ1酪氨酸磷酸化从而抑制生长信号转导是其治疗食管癌重要机制.

AIM： To investigate the signaling pathways involved in the action of Qigesan in treating esophageal cancer, the effects of Qigesan and its different individual components on tyrosine phosphorylation of PDGFR-PLC-γ1 were studied in primary cultured esophageal carcinoma cells. METHOOS： Cells from surgically resected human esophageal carcinoma specimens were primary cultured and treated with PDGF and the water extracts from Qigesan and its individual components. Total protein and tyrosine phosphorylated levels of PDGFR and PLC-γ1 were assessed by Western blotting. RESULTS： There was no difference in the levels of PDGFRβ and PLC-γ1 proteins before and after treatment of primary cultured esophageal carcinoma cells with water extracts from Qigesan and its separated components for 15 minutes and PDGF-BB for 4 minutes. The tyrosine phosphorylation levels of PDGFRβ and PLC-γ1 were markedly increased by stimulation with PDGFBB, and markedly decreased in cells treated with Qigesan and its separated components, which showed the best effects for promoting blood circulation group, while the second best was a whole prescription of Qigesan. CONCLUSION： Inhibiting growth signaling via inhibition of tyrosine phosphorylation of PDGFR and PLC-γ1 is an important mechanism in the treatment of esophageal cancer with Qigesan and its separated components.