去氧鬼臼毒素抑制肺癌NCI-H358细胞增殖和体外迁移的研究

Research on mechanisms of deoxypodophyllotoxin-induced inhibition of cell proliferation and migration in human lung cancer NCI-H358 cells

目的探讨去氧鬼臼毒素对人肺癌NCI-H358细胞增殖和体外迁移的影响,并初步探讨其作用机制。方法应用CCK-8法、流式细胞术、细胞划痕实验和DCFH-D法分别检测去氧鬼臼毒素对NCI-H358细胞增殖、周期分布、凋亡、体外迁移能力和细胞内活性氧(ROS)的影响,并通过Western blot检测去氧鬼臼毒素对细胞周期蛋白(Cyclin)B1、细胞分裂周期蛋白25同源蛋白(Cdc25c)、细胞周期蛋白依赖性激酶(CDK)1、半胱氨酸天冬氨酸蛋白酶(Caspase)-3、p53肿瘤蛋白、B淋巴细胞瘤(Bcl)-2和基质金属蛋白酶(MMP)9的表达水平,以及细胞外信号调节激酶(ERK)1/2、p38分裂原激活蛋白激酶(MAPK)和c-Jun氨基末端激酶(JNK)的磷酸化水平的影响。结果去氧鬼臼毒素能明显抑制肺癌NCI-H358细胞的生长,流式细胞结果显示其能诱导细胞停滞在G2/M和S期,诱导细胞凋亡和细胞ROS产生,同时细胞体外迁移能力也明显下调。Western blot结果显示,去氧鬼臼毒素能使Cyclin B1、Cdc25c、CDK1、Bcl-2和MMP9的蛋白表达明显下调,而Caspase-3和p53的表达明显上调,且ERK1/2、p38MAPK和JNK的磷酸化水平明显受到抑制。结论去氧鬼臼毒素可抑制肺癌NCI-H358细胞的增殖和体外迁移,是一种潜在的抗肿瘤药物。

Objective To investigate the effects and mechanism of deoxypodophyllotoxin on cell proliferation and mi？gration of human lung cancer NCI-H358 cells in vitro. Methods CCK-8 assay, flow cytometry assay, wound healing assay and DCFH-DA assay were used to detect the effects of deoxypodophyllotoxin on the proliferation, cells cycle, apoptosis, mi？gration and reactive oxygen species （ROS）. The protein expressions of Cyclin B1, Cdc25c, CDK1, Caspase-3, p53, Bcl-2, MMP9, ERK1/2, p38MAPK and JNK were measured by Western blot assay, respectively. Results Deoxypodophyllotoxin inhibited cell proliferation and reduced migration in human lung cancer NCI-H358 cells. Flow cytometry analysis showed that treatment with deoxypodophyllotoxin resulted in cell cycle G2/M and S phase arrest, cell apoptosis and ROS production. The result of Western blot assay showed that protein expressions of Cyclin B1, Cdc25c, CDK1, Bcl-2 and MMP9 were down-regulated while Caspase-3 and p53 were up-regulated. Moreover, Deoxypodophyllotoxin treatment decreased the phosphorylated levels of ERK1/2, p38MAPK and JNK obviously. Conclusion Deoxypodophyllotoxin could suppress the proliferation and migration of human lung cancer NCI-H358 cells in vitro, which is a potential anti-tumor drug.