白细胞介素35对非小细胞肺癌细胞增殖及凋亡的影响

Effect of Interleukin 35 on Proliferation and Apoptosis of Nonsmall Cell Lung Cancer Cells

[目的]评价沉默白细胞介素35(interleukin 35,IL-35)对非小细胞肺癌细胞增殖及凋亡的影响。[方法]运用短发卡RNA(short hairpin RNA,shRNA)干扰技术下调IL-35的表达,将转染shRNA的肺癌细胞分为shNC组和shIL35组,采用CCK-8法、流式细胞术分别检测细胞光密度(optical density,OD)值和凋亡率;实时荧光定量PCR(qPCR)和Western blot实验分别检测Cyclin D1、Bcl-2 mRNA和蛋白表达水平。[结果]利用shRNA下调A549、H1975细胞中IL-35的表达后,与shNC组相比,shIL35组OD值降低(t=8.452、8.064,P均<0.05),细胞凋亡率增加(t=5.745、4.334,P均<0.05);Cyclin D1、Bcl-2 mRNA和蛋白水平均表达下调(t=4.866、2.806、4.224、7.683,P均<0.05)。[结论]非小细胞肺癌细胞中IL-35的表达,可通过下调Cyclin D1、Bcl-2基因表达水平来抑制肺癌细胞增殖、促进其凋亡,为临床肺癌治疗提供新思路。

[Objective]To investigate the effect of silencing interleukin 35(IL-35)on proliferation and apoptosis of non-small cell lung cancer cells.[Methods]The short hairpin RNA(shRNA)interference technology was used to silence the expression of IL-35,and the lung cancer cell lines transfected with shRNA were divided into shNC group and shIL35 group.Then the optical density(OD)value and apoptosis rate of cells were detected respectively by CCK-8 method and flow cytometry,real-time fluorescent quantitative PCR(qPCR)and Western blot were used to detect the mRNA and protein expression of Cyclin D1 and Bcl-2,respectively.[Results]When silencing the expression of IL-35 with shRNA in A549 and H1975 cells,the OD value of shIL-35 group decreased(t=8.452,8.064,all P<0.05)and the apoptosis rate increased(t=5.745,4.334,all P<0.05)compared with the shNC group,and Cyclin D1,Bcl-2 mRNA and protein were down-regulated(t=4.866,2.806,4.224,7.683,all P<0.05).[Conclusion]The expression of IL-35 can inhibit the proliferation and promote the apoptosis of lung cancer cells by down-regulating Cyclin D1 and Bcl-2 proteins,it may provide some new ideas for clinical lung cancer gene therapy.