摘要
目的探究siRNA特异性降低肺癌细胞中巨噬细胞移动抑制因子(MIF)的表达量对A549细胞增殖、凋亡的影响以及作用机制。方法采用脂质体Lipofectamine2000将siRNA-MIF或siRNA-NC转入肺癌细胞中,实验分为3组:对照组、siRNA-NC组、siRNA-MIF组,噻唑蓝(MTT)检测细胞活力,流式细胞术检测细胞的凋亡率,免疫印迹试验(Western Blot)检测细胞中MIF、p65、磷酸化p65(p-p65)、细胞周期蛋白D1(cyclin D1)蛋白的表达水平。结果 siRNA-MIF组细胞中MIF蛋白的表达量显著低于对照组(P<0.05)。与对照组相比,siRNA-MIF组细胞的增殖显著下降(P<0.05),凋亡率显著增高(P<0.05)。siRNA-MIF组细胞中p65蛋白的表达量与对照组差异不显著(P>0.05),但p-p65(P<0.05)、cyclin D1(P<0.05)蛋白的含量显著降低。结论下调MIF的表达能有效抑制肺癌细胞的增殖,诱导其凋亡,其作用机制可能是通过调控NF-κB信号通路介导的下游靶基因的表达。
Objective To investigate the effect and mechanism on the proliferation and apoptosis by siRNA interferenced macrophage migration inhibitory factor( MIF) expression in lung cancer A549 cells. Methods siRNAMIF and siRNA-NC were transfected into lung cancer cells by lipofectamine 2000. The experiment was divided into 3 groups: the control group,the siRNA-NC group,and the siRNA-MIF group. The viability and apoptosis rate were assessed by MTT and flow cytometry. The expression of MIF,p65,phosphorylated p65( p-p65) and cyclin D1 were detected by Western blot. Results The expression of MIF protein was significantly lower in the siRNA-MIF group than in the control group( P〈0. 05). Compared with the control group,the proliferation were significantly decreased( P〈0. 05),and the apoptosis rate increased significantly in the siRNA-MIF group( P〈0. 05). The expression of p65 protein in the siRNA-MIF group was not significantly different from the control group( P〉0. 05),but p-p65 and cyclin D1 protein were significantly reduced. Conclusion The down-regulation of MIF expression can effectively inhibit the proliferation and induce apoptosis of lung cancer cells. The mechanism may be mediated through regulating the expression of downstream target genes by NF-κB signaling pathway.
作者
周勇
陈佳佳
蒋亚岚
ZHOU Yong;CHENG Jia-jia;JIANG Ya-lan(Department of Respiratory Medicine, Chongqing Rongchang District People’s Hospital, Chongqing, Sichuan 402460, Chin)
出处
《临床肺科杂志》
2018年第6期1001-1005,共5页
Journal of Clinical Pulmonary Medicine