Preliminary Mechanism Study Of ABCB6 Involved In The Metabolism Of Melanin

[Objective] To investigate preliminarily how ABCB6 interacts with key factors TYR,TYRP1, MC1 R and ISP during the melanin synthesis.[Methods] A wild type plasmid p Ds Red-wt-ABCB6 and a mutant type plasmid p Ds Red-L356P-ABCB6 were constructed using molecular biological technology.These plasmids and control plasmid were transfected A375 cells by Lipofectamine2000, respectively. The expression level of TYR, TYRP1, MC1 R, ISP m RNA and their protein level among these A375 cells were detected by Real-time PCR and Western blot, respectively.[Results] The p Ds Red-wt-ABCB6 plasmid and p Ds Red-L356P-ABCB6 plasmid were constructed successfully in this study. There were no statistically significant differences of the m RNA expression level of TYR, TYRP1, MC1 R, and ISP found between A375 cells transfected 32 hours from p Ds Red-wt-ABCB6 group,p Ds Red-L356P-ABCB6 group and p Ds Red-N1 group. No statistically significant differences of the protein level of TYR, TYRP1, MC1 R, and ISP were detected between A375 cells transfected 24 hours from p Ds Red-wt-ABCB6 group,p Ds Red-L356P-ABCB6 group and p Ds Red-N1 group. There were statistically significant lower of the protein level of TYRP1 discovered in A375 cells transfected32 hours or 40 hours from p Ds Red-L356P-ABCB6 group than p Ds Red-N1 group(P<0.05). However, similar findings were not found in A375 cells from p Ds Red-wt-ABCB6 group. Meanwhile, no statistically significant differences of the protein level of TYR, MC1 R, and ISP were detected between A375 cells transfected32 hours or 40 hours from p Ds Red-wt-ABCB6 group, p Ds Red-L356P- ABCB6 group and p Ds Red-N1 group.[Conclusions] Mutant ABCB6 had little impact on the m RNA levels of TYR, TYRP1,MC1 R, ISP in A375 cells. Most likely, ABCB6 interacted on TYRP1 protein in the process of melanin metabolism. Its molecular mechanism need further research.