c-Jun氨基末端激酶1和2在角质形成细胞系Hacat细胞中的作用

Role of c-Jun amino-terminal kinase 1 and 2 in human keratinocyte cell line Hacat

目的研究同工酶JNK1和JNK2在Hacat细胞中的功能差异,探讨其在皮肤肿瘤治疗中的潜在价值。方法分别将JNK1和JNK2的siRNA导入Hacat细胞,通过Western Blot技术在蛋白质水平对其敲低效果进行鉴定。CCK-8法检测细胞增殖曲线,流式细胞术检测细胞周期,不同剂量的Taxol进行凋亡诱导实验,分析正常Hacat细胞与JNK1/2敲低细胞的区别。结果细胞增殖实验表明,JNK1或JNK2敲低均可导致增殖变缓,JNK1敲低效果更明显。细胞周期结果显示,JNK1敲低可导致S期细胞减少,G2/M期升高。Taxol凋亡诱导实验表明,JNK2敲低组凋亡显著增加,而对照组和JNK1敲低组凋亡变化不明显。结论在细胞增殖方面,JNK1作用强于JNK2;而在抗凋亡方面,JNK2作用强于JNK1。

Objective To study the different roles of knocked – down c-Jun amino-terminal kinase 1（JNK1） and c-Jun aminoterminal kinase 2（JNK2） in expression of Hacat cells.Methods JNK1 and JNK2 siRNA were transfected into Hacat cells.The expression level of Hacat cells was measured by Western blot.The proliferation of Hacat cells was detected by CCK-8 assay.The cell cycle of Hacat cells was assayed by flow cytometry.The apoptosis of Hacat cells was induced by different doses of Taxol.Different expressions of normal and JNK1/2 knocked-down Hacat cells were analyzed.Results The knocked-down JNK1 and JNK2,especially JNK1,could reduce the proliferation of Hacat cells.The knocked-down JNK1 could decrease the number of Hacat cells at S phase and increase the number of Hacat cells at G2/M phase.The apoptosis of Hacat cells increased significantly in knocked-down JNK2.However,no significant difference was found in normal and JNK1 knocked-down Hacat cells.Conclusion The effect of JNK1 is better than that of JNK2 on proliferation of Hacat cells,whereas the effect of JNK2 is stronger than that of JNK1 on apoptosis of Hacat cells.