MC1R对白癜风黑色素细胞增殖和凋亡的影响研究

Effect of MC1R on proliferation and apoptosis of vitiligo melanocytes

目的探讨黑素皮质受体1(MC1R)对白癜风黑色素细胞的作用和相关调控机制。方法体外培养人表皮黑色素细胞系PIG1和白癜风黑色素细胞系PIG3V,分别用MC1R siRNA和MC1R过表达载体处理,qRT-PCR和Western Blot检测各组细胞中MC1R的表达,MTT检测细胞增殖改变,流式细胞术检测细胞凋亡和细胞周期改变,活性氧试剂盒检测活性氧(ROS)的产生情况,Western Blot检测p38促分裂原活化蛋白激酶(MAPK)和c-Jun氨基末端激酶(JNK)、Caspase 3、Bax和Bcl-2、p53、p21和Cyclin D1表达情况。结果敲减MC1R表达后MC1R表达下降,显著抑制黑色素细胞增殖,促进细胞凋亡,G2/M期细胞比例增高,ROS水平增加,Bcl-2、Cyclin D1蛋白降低,p-p38 MAPK、JNK、Caspase 3、Bax、p53和p21蛋白水平升高(P<0.05);而过表达MC1R后逆转上述改变(P<0.05)。结论调控MC1R表达可引起黑色素细胞生物学行为改变,可能参与白癜风黑色素细胞的病变过程。

Objective To explore the effect of melanocortin 1 receptor(MC1R)on vitiligo melanocytes and its related regulatory mechanism.Methods Human epidermal melanocyte cell line PIG1 and vitiligo melanocyte cell line PIG3V were cultured in vitro,treated with MC1R siRNA and MC1R overexpression vector,respectively.qRT-PCR and Western Blot were used to test the expression of MC1R,MTT was used to test the change of proliferation,flow cytometry was used to test the changes of apoptosis and cell cycle,reactive oxygen species kit was used to test the production of reactive oxygen species(ROS),Western Blot was used to test the expressions of p38 mitogen-activated protein kinase(MAPK)and c-Jun N-terminal kinase(JNK),Caspase 3,Bax and Bcl-2,p53,p21,and Cyclin D1.Results After knockdown of MC1R expression,the expression of MC1R decreased,leading to significantly inhibited proliferation of melanocytes,promoted apoptosis,increased proportion of G2/M cells,increased ROS levels,decreased Bcl-2 and Cyclin D1 proteins,and increased levels of p-p38 MAPK,JNK,Caspase 3,Bax,p53,and p21 proteins(P<0.05).However,overexpression of MC1R reversed the above changes(P<0.05).Conclusion The regulation of MC1R expression can cause changes in the biological behavior of melanocytes,which may be involved in the pathological process of vitiligo melanocytes.