摘要
目的:探讨胰岛素样生长因子结合蛋白家族4(IGFBP-4)在灯盏花素(BVP)抑制肺癌A549细胞增殖中的作用。方法:培养肺癌A549细胞,用0、25、50和100 μmol/L BVP处理A549细胞48 h后,用MTT方法检测细胞存活率,台盼蓝染色法检测死亡细胞比例,Annexin V/Propidium iodide(PI)双染法检测细胞凋亡,Western blot、qRT-PCR和ELISA方法检测Bax、Bcl-2及IGFBP-4的表达和分泌情况。建立IGFBP-4高表达和低表达A549细胞,经100 μmol/L BVP处理48 h后,用MTT方法检测细胞存活率,Western blot检测Bax、Bcl-2的表达。结果:25、50和100 μmol/L BVP显著减低细胞存活,增加细胞凋亡,上调Bax表达,下调Bcl-2表达。25、50和100 μmol/L BVP上调细胞内IGFBP-4 m RNA和蛋白表达以及增加培养基中IGFBP-4含量。IGFBP-4高表达降低A549细胞存活率,增加BVP抗A549细胞增殖作用,IGFBP-4低表达增加细胞存活率,部分降低BVP抗A549细胞增殖作用。结论:BVP具有抗A549细胞增殖作用,其机制可能与其上调IGFBP-4表达有关。
Objective： To investigate the role of IGFBP-4 in breviscapine （BVP） inhibited lung cancer A549 cells growth. Methods： A549 cells were treated with BVP 0, 25, 50 and 100 μmol/L BVP for 48 h. Adv-IGFBP-4-OE？ DNA, EGFP and Adv-KD？ Easy RNAi EGFP were used to establish IGFBP-4 over- or down- expres-sion A549 cells. MTT assay was used to detect cell survival rate, Trypan blue staining was applied to analyze cell death rate, Annexin V/Propidium iodide （PI） double staining was used to test cell apoptosis. Western blot, RT-qPCR and ELISA were used to detect the expression Bax, Bcl-2, the protein and mRNA expression and secretion of IGFBP-4. Results： The viability decreased, but apoptosis rate increased signifcantly in A549 cells with 25, 50 and 100 μmol/L BVP treatment. BVP also increased Bax but reduced Bcl-2 expression. The mRNA and protein expression of IGFBP-4 and the secretion of IGFBP-4 increased with BVP treatment for 48 h. The over-expres-sion of IGFBP-4 decreased the survival rate of A549 cells, and enhanced the growth inhibition by 100 μmol/L BVP. The down-expression of IGFBP-4 increased the cell survival rate, and alleviated the growth inhibition by 100 μmol/L BVP. Conclusion： BVP has an inhibitory effect on A549 growth, which might be related to overex-pression of IGFBP-4.
作者
孙蕾
李伟文
黎媛
蓝秀
吕祝庆
Sun Lei;Li Weiwen;Li Yuan;Lan Xiu;LYu Zhuqing(Department of Respiratory Medicine,Lishui Hospital of Zhejiang university,the Fifth Affliated Hospital of Wenzhou Medical University,Lishui Municipal Central Hospital,Lishui,32300)
出处
《温州医科大学学报》
CAS
2018年第11期813-817,共5页
Journal of Wenzhou Medical University
基金
浙江省科技厅公益性技术应用研究计划项目(2016c37100)
