虎杖苷对肺癌A549细胞增殖和侵袭的抑制作用及机制探讨

目的 探讨虎杖苷对肺癌A549细胞增殖和侵袭的抑制作用，并探讨其机制。方法 以肺癌A549细胞为研究对象，以不同浓度虎杖苷（0、10、20、50、100、200 mg/L）处理24、48和72 h后，四氮唑盐（MTS）实验检测虎杖苷对肺癌A549细胞体外增殖的作用，以确定后续实验浓度；经不同浓度虎杖苷（0、10、20 mg/L）处理48 h后，Transwell实验检测虎杖苷对肺癌A549细胞体外侵袭的作用；Western blot和Real-time PCR检测基质金属蛋白酶2/9（MMP-2/9）、整合素β4（integrin β4）、Toll样受体3（TLR3）和分化抗原44（CD44）的表达以及丝氨酸/苏氨酸激酶磷酸化（p-AKT）水平；酶联免疫吸附试验（ELISA）检测培养基中转化生长因子β（TGF-β）和肿瘤坏死因子-α（TNF-α）含量；报告基因技术检测核因子（NF）-κB启动子活性。结果 在0、10、20、50、100、200 mg/L浓度范围内，虎杖苷处理肺癌A549细胞后，细胞体外增殖率显著下调（以10 mg/L和20 mg/L处理48 h作为后续实验的药物浓度和时间）。虎杖苷处理48 h后，肿瘤细胞体外侵袭能力显著下降，TGF-β和TNF-α含量显著下调，MMP-2/9、integrin β4、TLR3、CD44、p-AKT蛋白及mRNA水平以及NF-κB启动子活性显著下调（均P＜0.05）。结论 通过调控肿瘤相关基因表达和ART/NF-κB信号通路活性，虎杖苷可显著抑制肺癌A549细胞增殖和侵袭。

The inhibitive effect of polydatin on the proliferation and invasion in lung cancer A549 cells and its mechanism

Objective To explore the inhibitory effect of polydatin on the proliferation and invasion in lung cancer A549 cells and its mechanism. Methods The lung cancer A549 cells were treated with different concentrations of polydatin (0, 10, 20, 50, 100 and 200 mg / L) for 24, 48 and 72 hours. The proliferation of A549 cells was measured by tetrazolium nitrogen salt (MTS) assay to determine the concentration of subsequent experiments (10 mg/L and 20 mg/L for 48h). With different concentrations of polydatin (0, 10 and 20 mg/L) for 48 hours, the invasion of A549 cells was measured by Transwell assay. The expression levels of matrix metalloprotinase (MMP)-2/9, integrin β4, Toll-like receptor (TLR3), cluster of differentiation (CD44) and phosphorylation of serine/threonine kinase (p-AKT) were measured by Western blotting and real-time PCR assay. The content of transforming growth factor (TGF) - β and tumor necrosis factor (TNF) - α in culture medium were measured by ELISA assay. The activity of nuclear factor (NF) - κB was measured by report gene assay.Results In the concentration ranges of 0, 10, 20, 50, 100 and 200 mg/L polydatin for 24, 48 and 72 h, the cell proliferation was decreased, and 10 mg/L and 20 mg/L of polydatin treatment for 48 h were used for following experiments. After treatment with polydatin for 48 h, the invasion was decreased, the contents of TGF-β and TNF-α were decreased, and the expression levels of MMP-2 / 9, integrin β4, TLR3, CD44, p-AKT and the activity of NF - κB were also decreased (P＜0.05).Conclusion Polydatin can inhibit the proliferation and invasion of lung cancer A549 cells by regulating the expression of tumor reated gene and the activity of AKT/NF-κB signal pathway.