Rab23分子对中波紫外线诱导人黑素细胞系PIG1细胞黑素合成的调控效应

Role of Rab23 in melanogenesis induced by ultraviolet radiation B in human melanocyte cell line PIG1

目的探究Rab23是否参与调控中波紫外线(ultraviolet B,UVB)诱导的黑素细胞黑素合成。方法分别用25 mJ/cm^2、50 mJ/cm^2、100 mJ/cm^2 UVB照射人黑素细胞系PIG1细胞,Western印迹检测Rab23及小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)的蛋白表达水平。用50 mJ/cm^2 UVB照射正常人表皮黑素细胞(PIG1)后,分别于0.5 h、1 h、2h、6 h、12 h提取细胞蛋白,Western印迹检测Rab23及黑素合成相关蛋白MITF、gp100的表达水平。在PIG1细胞中构建Rab23的RNA干扰细胞系(siRNA),使用50 mJ/cm^2 UVB处理细胞,继续培养6 h后采用Western印迹检测黑素合成相关蛋白MITF、gp100的表达水平。结果 Rab23及MITF表达量随UVB剂量增大而增大;50 mJ/cm^2 UVB照射黑素细胞,培养时间越长,Rab23表达水平越高,MITF和gp100表达水平也越高;使用50 mJ/cm^2 UVB照射Rab23的RNA干扰细胞系继续培养6 h后,细胞中的MITF和gp100表达水平较Rab23正常表达组明显降低。结论 UVB可剂量和时间依赖性上调Rab23及黑素合成相关蛋白表达水平,下调Rab23表达可降低UVB诱导的MITF和gp100表达。

Objective To explore the role of Rab23 in melanogenesis induced by UVB in human melanocytes. Methods The human melanocyte PIG1 cells were irradiated respectively with UVB at 25 mJ/cm^2, 50 mJ/cm^2 and 100mJ/cm^2, and the western blot was performed to determine the expression of Rab23 and MITF in PIG1 cells. The protein of PIG1 cells treated with UVB at 50 mJ/cm^2 was extracted respectively after 0.5 h,1 h,2 h,6 h and 12 h,then the western blot was conducted to measure the expression of Rab23, MITF and gp100. PIG1 cells transfected by siRNA targeting Rab23 were treated with UVB at 50 mJ/cm^2, and its protein was extracted after 6 h, then the western blot was conducted to measure the expression of Rab23, MITF and gp100. Results UVB irradiation promoted the expression level of Rab23, MITF and gp100 in PIG1 cells in a positive dose-dependent and time-dependent manner, while the Rab23 inhibition in PIG1 cells decreased the expression level of MITF and gp100 induced by UVB of 50 mJ/cm^2 for 6 hours irradiation. Conclusion Rab23 is involved in the melanogenesis induced by irradiation with UVB, and the downregulation of Rab23 will decrease MITF and gp100 induced by UVB-stimulated .