消岩汤通过WNT通路抑制肺腺癌A549细胞增殖、促进细胞凋亡的机制研究

Study on the mechanism of Xiaoyan Decoction inhibiting proliferation and promoting apoptosis of lung adenocarcinoma A549 cells through WNT pathway

[目的]探讨扶正祛瘀解毒法经典用药消岩汤抑制人肺腺癌A549细胞增殖、促进细胞凋亡的可能机制。[方法]体外培养A549细胞,乳酸脱氢酶(LDH)、CCK8、流式细胞术检测对照组、消岩汤组、消岩汤加WNT通路激动剂组、WNT通路抑制剂XAV939组、单独WNT通路激动剂组细胞毒性、存活率和凋亡率。通过聚合酶链式反应(RT-PCR)测定对照组、消岩汤组和WNT通路抑制剂XAV939组中WNT通路相关基因β-catenin、c-myc,凋亡相关基因Bcl-2、Caspase-3表达情况;通过荧光素酶报告基因检测反应WNT通路活性。[结果]消岩汤可以抑制A549细胞的增殖,促进细胞凋亡,而消岩汤加WNT通路激动剂组可以减弱以上作用;同时消岩汤可以显著降低A549细胞中β-catenin、c-myc的表达水平,抑制WNT通路活性,并通过降低Bcl-2表达、增强Caspase-3表达来促进了肿瘤细胞凋亡。[结论]扶正祛瘀解毒法消岩汤可以抑制A549细胞的增殖、促进凋亡,其作用机制可能是抑制WNT通路活性。

[Objective]To investigate the possible mechanism of“Xiaoyan Decoction”classical medicine of strengthening the body resistance,removing blood stasis and detoxifying method,to inhibit A549 cell proliferation and promote cell apoptosis in human lung adenocarcinoma.[Methods]A549 cells were cultured in vitro,and the cytotoxicity,survival rate and apoptosis rate of the control group,Xiaoyan Decoction group,Xiaoyan Decoction plus WNT pathway agonist group,XAV939 group and WNT pathway agonist group were detected by LDH,CCK8 and flow cytometry.The expression levels ofβ-catenin,c-myc,Bcl-2 and Caspase-3 related genes in WNT pathway were determined by PCR.WNT pathway activity was detected by luciferase reporter gene assay in control group,Xiaoyan Decoction group and WNT pathway inhibitor XAV939 group.[Results]Xiaoyan Decoction can inhibit the proliferation of A549 cells and promote apoptosis,while Xiaoyan Decoction plus WNT pathway agonist group can weaken the above effect,meanwhile,Xiaoyan Decoction can significantly reduce the expression levels ofβ-catenin and c-myc in A549 cells in a dose-dependent manner,inhibit the activity of WNT pathway,and promote the apoptosis of tumor cells by reducing the expression of Bcl-2 and enhancing the expression of caspase-3.[Conclusion]Xiaoyan Decoction may inhibit the proliferation and promote apoptosis of A549 cells by inhibiting the WNT signaling pathway.