MGluR6 Regulates Keratinocyte Phagocytosis By Modulating Activity Of CaM KII/REK/MLC Signaling Pathway

Objective: To explorer the molecular mechanism of metabotropic glutamate receptor 6 regulates keratinocyte phagocytosis.Methods: 1.The cell culture of h?man keratinocyte cell lines,HaCaT cells.2.Western blotting analysis of the expression of m GluR6 in HaCaT cells.3.Knockdown of m GluR6 in HaCaT cells using GFP-tagged lentiviral vectors sh RNA for m GluR6 and a negative control(NC).4.The knockdown efficiency of m GluR6 in HaCaT cells was investigated by western blot.5.The distribution of cytoskeleton F-actin in each group was visualized by scanning laser confocal microscopy.6.The expression of pERK and ERK in each group was analyzed by western blot.7.The expression of pCaM KII and CaM KII in each group was analyzed by western blot.8.The expression of p MLC and MLC in each group was analyzed by western blot.Results: 1.The HaCaT cells express m GluR6.2.The stably knockdown of m GluR6 HaCaT cells were established successfully,and the knockdown efficiency was nearly 80%.3.The HaCaTm GuR-l6 cells represented reorganization of F-actin cytoskeleton,compared with control cells.4.The expression of pERK in HaCaT-m Glu R6 cells was significantly decreased,compared with control cells.The ratio of pERK and ERK was decreased.5.The expression of pCaM KII in HaCaT-m Glu R6 cells was significantly decreased,compared with control cells.The ratio of pCaM KII and CaM KII was decreased.6.The expression of pMLC in HaCaT-m Glu R6 cells was significantly decreased,compared with control cells.The ratio of p MLC and MLC was decreased.Conclusion: 1.The human keratinocytes express m GluR6.2.mGluR6 involves in the regulation of keratinocyte cytoskeleton F-actin distribution.3.mGluR6 regulates keratinocyte phagocytosis through CaM KII/ERK/MLC signaling pathway.