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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.
OBJECTIVE: To observe the effects of Qige San and its activating blood and resolving phlegm on tyrosine phosphorylation of platelet-derived growth factor receptor (PDGFR) and phospholipase C-γ1 (PLC-γ1) in primary cultured esophageal cancer cells In order to investigate the mechanism of Qige Powder’s treatment of esophageal cancer.Methods: Esophageal epithelial cells were harvested from surgically resected primary esophageal cancer tissues for primary culture, and PDGF and Qige Powder were added to two groups PDGF-BB was used to measure the expression of PDGFR-PLC-γ1 and the level of tyrosine phosphorylation.Results: PDGF-BB stimulated for 4 min and Kaige powder and its disassembled recipe for 15 min PDGF-BB stimulated tyrosine phosphorylation of PDGFRβ and PLC-γ1 protein significantly, while QGD and its disassembled recipe decreased to some extent The phosphorylation level of the protein was the highest in the blood-activating group, followed by the other groups.Conclusion: The pathological changes of esophageal cancer are related to the enhancement of tyrosine phosphorylation of PDGFR-PLC-γ1 protein, The phosphorylation of PLC-γ1 tyrosine which inhibits growth signal transduction is its treatment of the esophagus Cancer important mechanism.