基于JAK2/STAT3/VEGF通路探讨周氏固金消瘤汤抗人肺腺癌A549细胞的机制研究

目的   研究周氏固金消瘤汤(ZSGJXLT)对人肺腺癌A549增殖、凋亡和血管生成的影响及其机制。方法   通过CCK-8法检测ZSGJXLT对细胞活性的影响；Annexin V-FITC/PI流式细胞术检测细胞凋亡；体外血管形成实验观察血管生成情况；Western blot法检测Bcl-2、Bax、VEGF、JAK2、p-JAK2、STAT3及p-STAT3表达；构建小鼠A549肺癌皮下移植瘤模型，并给予不同浓度ZSGJXLT治疗，21 d后处死小鼠称取瘤质量，计算抑瘤率。结果   ZSGJXLT显著抑制A549的活性(P < 0.01)，具有时间及浓度依赖性；ZSGJXLT诱导A549细胞凋亡(P < 0.05~0.01)，具有浓度依赖性；ZSGJXLT显著抑制人脐静脉内皮细胞(HUVEC)体外小管形成(P < 0.01)，呈浓度依赖性；Western blot结果显示ZSGJXLT能引起Bax/Bcl-2表达明显上调(P < 0.05~0.01)，VEGF、p-STAT3/STAT3和p-JAK2/JAK2的表达明显下调(P < 0.01)；小鼠体内实验结果表明，与模型组比较，不同浓度ZSGJXLT(0.25、1、4 g/kg)均可显著抑制肿瘤生长(P < 0.05~0.01)，抑瘤率分别为21.66%、35.52%和48.24%。结论   周氏固金消瘤汤可以通过影响JAK2/STAT3/VEGF通路的活化，发挥抑制肿瘤细胞增殖，促进肿瘤细胞凋亡，抗血管生成等多途径抗人肺腺癌A549的作用。为开发新型抗肺癌中药复方ZSGJXLT提供了实验依据。 

Study on the Mechanism of Zhoushi Gujin Xiaoliu Tang Against Human Lung Adenocarcinoma A549 Cells Based on JAK2/STAT3/VEGF Pathway

OBJECTIVE To investigate the effect and mechanism of Zhoushi Gujin Xiaoliu Tang(ZSGJXLT)on proliferation,apoptosis and angiogenesis of human lung adenocarcinoma A549 cells.METHODS The activity of A549 cells was detected by CCK-8.The apoptosis of A549 cells was observed by the Annexin-V-FITC/PI flow cytometry.The cell angiogenesis ability was evaluated by human umbilical vein endothelial cell(HUVEC)angiogenesis assay in vitro.The expression of Bcl-2,Bax,VEGF,JAK2,p-JAK2,STAT3 and p-STAT3 proteins were detected by Western blot.The A549 lung cancer xenograft model was constructed and treated with ZSGJXLT,which was divided into different concentrations.After 21 days,the mice were sacrificed to weigh the tumors and the tumor inhibitory rate was calculated.RESULTS The proliferation of A549 were apparently inhibited by ZSGJXLT in a concentration and time-dependent manner(P<0.01).ZSGJXLT also induced A549 cell apoptosis significantly in a concentration-dependent manner(P<0.05,P<0.01).In addition,the number of annular vascular structure was decreased by ZSGJXLT(P<0.01).The result of Western blot showed that ZSGJXLT promoted the up-regulation of Bax/Bcl-2(P<0.05,P<0.01)and inhibited the expression of VEGF,p-STAT3/STAT3 and p-JAK2/JAK2(P<0.05).In vivo,ZSGJXLT(0.25,1,4 g/kg)also statically inhibited the growth of lung cancer xenograft in nude mice model in a concentration-dependent manner(P<0.05,P<0.01),with tumor inhibitory rates of 21.66%,35.52% and 48.24%,respectively.CONCLUSION ZSGJXLT inhibits the activation of JAK2/STAT3/VEGF signaling pathway,thereby inhibiting tumor cell proliferation,promoting tumor cell apoptosis,anti-tumor angiogenesis and other ways to play anti-tumor effects.This research provides experimental evidence of ZSGJXLT for lung cancer.