丝氨酸/苏氨酸蛋白激酶对肝癌细胞凋亡的促进作用

HIPK1 induced hepatoma cell apoptosis

目的评价丝氨酸/苏氨酸蛋白激酶(HIPK1)促进肿瘤坏死因子(TNF)-α诱导的JNK/p38激活及其对肝癌细胞凋亡的影响。方法Hepa1-6细胞,分别感染重组腺病毒rAd-Lacz-V5(107PFU/孔)和rAd-myc-HIPK1(107PFU/孔)48h,荧光显微镜检测Lacz和HIPK1的蛋白表达,TNF-α处理细胞后Western blot检测JNK/p38激活情况;HIPK1的RNA干扰检测HIPK1在JNK/p38激活中的作用;TNF-α处理病毒感染的Hepa1-6细胞24h,DAPI染色荧光显微镜下了解细胞凋亡情况。结果荧光显微镜下见Lacz和HIPK1蛋白在Hepa1-6细胞中表达效率高;HIPK1组中TNF-α介导的磷酸化型JNK/p38的表达明显高于Lacz组,相反HIPK1的RNA干扰组中TNF-α介导的磷酸化型JNK/p38的表达明显低于对照组;细胞核形态学检测HIPK1组中细胞凋亡率为(30.72±5.67)%,而Lacz组为(9.82±3.41)%,两组间差异有统计学意义(P<0.01)。结论HIPK1促进TNF-α介导的JNK/p38的激活并诱导肝癌细胞Hepa1-6凋亡。

Objectibve To assess the role of HIPK1 on TNF-α- induced JNK/p38 activation and hepatoma cell apoptosis. Methods Hepal-6 cells were infected with rAd-Lacz-V5 （101 PFU/well） and rAd-myc-HIPK1 （101 PFU/weU）, respectively. Forty-eight h later, the expression of protein Laez and HIPK1 was detected by immunoflouresenee. After the cells were treated with TNF-α, Western blotting was used to detect the phosphorylation of JNK/p38. Moreover, HIPK1 was knockdown by RNA interference in Hepa1-6 cells and the cells were treated with TNF-α, and phosphorylation of JNK/p38 in those cells was checked by Western blotting. The morphology of nuclei in each group was revealed by DIPA staining. Results Over-expression of HIPK1 increased the TNF-α-indueed JNK/p38 activation, whereas small interference RNA of HIPK1 decreased the TNF-α-indueed JNK/p38 activation. Consistently, the percentage of apoptotie cells in HIPK1 group was （30.72 ± 5.67） %, but （9.82±3.41） % in Lacz group （P〈0.01）. Conclusion HIPK1 enhances the TNF-α-indueed JNK/p38 activation and promotes the apoptosis of Hepal-6 hepatoma cells.