Tang Yinhong,Yang Ke,Yang Tian,Ye Jixing,Liu Peng,Lian Xiaohua,Guo Haiying,Peng Huiming.SFRP5 suppresses melanin formation of melanocytes through Wnt3a inhibition[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(03):228-231.
Tang Yinhong,Yang Ke,Yang Tian,Ye Jixing,Liu Peng,Lian Xiaohua,Guo Haiying,Peng Huiming.SFRP5 suppresses melanin formation of melanocytes through Wnt3a inhibition[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(03):228-231.DOI:
SFRP5 suppresses melanin formation of melanocytes through Wnt3a inhibition
Objective To explore the effects of secreted Frizzled-related protein 5 (sFRP5) inhibiting Wnt3a on the melanin formation of melanocytes. Methods With immortalized melanocytes as model and adenoviral vectors
we set up 3 experimental groups
melanocytes transfected with Ad-GFP (Ad-GFP group)
melanocytes transfected with Ad-Wnt3a (Ad-Wnt3a group) and melanocytes transfected with Ad-Wnt3a+Ad-sFRP5 (Ad-Wnt3a+Ad-sFRP5 group) to do the following experiments
including L-dopa tyrosinase activity assay
Masson-Fontanas silver staining
for melanin formation detection
RT-PCR for testing expression of tyrosinase-related protein 1 (TRP-1) and tyrosinase (TYR)
and Western blotting for detecting the protein levels of TRP-1 and TYR in iMC23 cells. Results L-Dopa tyrosinase activity assay showed an obvious decrease in tyrosinase activity in the Ad-Wnt3a+Ad-sFRP5 group as compared to Ad-Wnt3a group (
P
0.05). Masson-Fontanas silver staining found that melanin in the Ad-Wnt3a+Ad-sFRP5 group was significantly lower than that in the Ad-Wnt3a group. RT-PCR results showed that the mRNA expression levels of TRP-1 and TYR were down-regulated in the Ad-Wnt3a+Ad-sFRP5 group (
P
0.05). Moreover
the protein levels of TRP-1 and TYR showed similar changes as proven by Western blotting (
P
0.05). Conclusion SFRP5 can suppress the melanin formation of melanocytes through Wnt3a inhibition.