玉屏风散含药血清对肺癌A549细胞及STAT3信号通路的影响

Effects of serum containing Yupingfeng Powder on A549 cells and STAT3 signaling pathway of lung cancer

目的 探讨玉屏风散含药血清对肺癌A549细胞增殖、周期、凋亡、侵袭及信号转导和转录激活因子3(STAT3)信号通路的影响。方法 将体外培养的A549细胞分为对照组(给予空白血清)和低剂量、中剂量、高剂量组(给予低剂量、中剂量、高剂量玉屏风散含药血清),采用噻唑蓝(MTT)法检测A549细胞增殖,流式细胞仪检测A549细胞周期和凋亡,Transwell小室检测A549细胞侵袭,免疫印迹法(Western blot)检测A549细胞中STAT3、磷酸化(p)-STAT3、细胞周期蛋白D1(Cyclin D1)、B淋巴细胞瘤-2基因(Bcl-2)和基质金属蛋白酶9(MMP-9)蛋白表达。结果 与对照组比较,低剂量、中剂量、高剂量玉屏风散含药血清刺激后A549细胞增殖明显受到抑制(P <0. 05),且呈一定的时间-剂量依赖性(P <0. 05)。与对照组比较,低剂量、中剂量、高剂量组细胞凋亡率和G2/M期细胞所占百分比均明显升高(P <0. 05),而穿膜细胞数、S期细胞所占百分比和STAT3磷酸化水平以及Cyclin D1、Bcl-2、MMP-9蛋白表达水平均明显降低(P <0. 05),且呈一定的剂量依赖性(P <0. 05)。结论 玉屏风散含药血清可抑制肺癌A549细胞增殖、侵袭并诱导细胞周期阻滞和凋亡,其作用机制可能与抑制STAT3信号通路活化有关。

Objective To investigate the effects of Yupingfeng Powder-containing serum on the proliferation,cycle,apoptosis,invasion,signal transducer and activator of transcription 3(STAT3)signal pathway of lung cancer A549 cells.Methods A549 cells cultured in vitro were divided into control group(given blank serum)and low,medium and high dose groups(given low,medium and high dose Yupingfeng powder—containing serum);the proliferation of A549 cells was detected by Methyl Thiazolyl Tetrazolium(MTT);flow cytometry was used to detect A549 cell cycle and apoptosis,the invasion of A549 cells was detected by Trans well cell,and Western blot was used to detect the expressions of STAT3,phosphorylation(p)—STAT3,CyclinDl,B-lymphoma-2 gene(Bel-2)and matrix metalloproteinase-9(MMP-9)proteins in A549 cells.Results Compared with the control group,the proliferation of A549 cells was significantly inhibited by the stimulation of low,medium and high doses of Yupingfeng Powder-containing serum(P<0.05),which is time-dose dependent(P<0.05).Compared with those in the control group,the apoptosis rate and the percentage of G2/M phase cells in the low,middle and high dose groups were significantly higher(P<0.05);however,the number of transmembrane cells,the percentage of S phase cells,the phosphorylation level of STAT3 and the expression levels of CyclinDl,Bel-2 and MMP-9 proteins were significantly lower(P<0.05),which is dose-dependent(P<0.05).Conclusion Yupingfeng Powder-containing serum can inhibit the proliferation and invasion of lung cancer A549 cells and induce cell cycle arrest and apoptosis.The mechanism may be related to the inhibition of STAT3 signaling pathway activation.