白细胞介素-8沉默对喉鳞状细胞癌细胞增殖和凋亡的影响及机制研究

Effect of interleukin-8 silencing on proliferation and apoptosis of laryngeal squamous carcinoma andits mechanism

目的探讨白细胞介素-8(IL-8)沉默对喉鳞状细胞癌细胞增殖和凋亡的影响及相关机制。方法将IL-8小干扰RNA(siRNA)和Lipofectamine 2000转染试剂转染至喉鳞状细胞癌Hep-2细胞作为实验组和NC组仅转染Lipofectamine 2000转染试剂的细胞作为,未做处理的细胞作为空白组。采用噻唑蓝(MTT)法检测细胞增殖能力,流式细胞仪检测细胞凋亡情况,蛋白质印迹法(Western blot)检测IL-8、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、cleaved caspase 3、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(B AX)的相对表达量。结果实验组喉鳞状细胞癌Hep-2细胞中IL-8蛋白的相对表达量明显低于NC组和空白组细胞,OD值明显低于NC组和空白组细胞(P<0.01)。实验组喉鳞状细胞癌Hep-2细胞的凋亡率明显高于空白组和NC组细胞(P<0.01)。实验组Hep-2细胞中p-AKT和抑制凋亡蛋白Bcl-2的相对表达量均低于空白组和NC组细胞,而促凋亡蛋白B AX和cleaved caspase 3的相对表达量均高于空白组和NC组细胞(P<0.05)。3组Hep-2细胞AKT蛋白的相对表达量比较,差异均无统计学意义(P>0.05)。结论IL-8可通过对磷酸肌醇3-激酶(PI3K)/AKT相关信号通路的调控,抑制喉鳞状细胞癌Hep-2细胞的增殖能力,并促进其凋亡,为喉鳞状细胞癌的诊断和治疗提供了新的靶点及可能的治疗策略。

Objective To investigate the effect of interleukin-8(IL-8)silencing on proliferation and apoptosis of laryngeal squamous carcinoma and related mechanism.Method IL-8 was silenced by small interfering RNA(siRNA)via lipofectamine 2000 transfection reagentin Hep-2 cells andset as experiment group while Hep-2 cells with vector transfection were set as NC group(only add Lipofectamine 2000 transfection reagent)and Hep-2 cells without any treatment were set as blank group.Methyl-thiazolyldiphenyl-tetrazolium bromide(MTT)assay was employed to explore proliferation of Hep-2 cells.Flow cytometry assay was used to detect the apoptosis of Hep-2 cells and the relative protein expression of IL-8,protein kinase B(AKT),phosphorylated-AKT(p-AKT),cleaved caspase 3,B cell lymphoma/leukemia-2(Bcl-2)and Bcl-2 associated X protein(BAX)were detected by Western Blot.Result The relative expression of IL-8,together with the optical density(OD)value,in laryngeal squamous cancer Hep-2 cells from experiment group was significantly lower than cells in NC group and blank group with statistic difference(P<0.01).The apoptosis rate of laryngeal squamous cancer Hep-2 cells in experiment group was significant higher than that in NC group or blank group(P<0.01).The relative p-AKT protein and apoptosis inhibition Bcl-2 protein levels were lower while pro-apoptotic protein BAX and cleaved caspase3 relative levels in experiment group were higher in experiment group compared to NC group or blank group with statistic significant difference(P<0.05).However,there was no significant difference of AKT protein level among experiment group,NC group and blank group(P>0.05).Conclusion IL-8 could inhibit the proliferation of laryngeal squamous cancer Hep-2 cells and promote the apoptosis of cancer cellsvia modulating phosphatidylinositide 3-kinase(PI3 K)/AKT signaling pathway.This finding provides a new target and potential treatment strategy for the diagnosis and treatment for laryngeal carcinoma cells.