Mechanish Reseacher On Effects On Cell Structure Of KRT9R163W Mutant In Epidermolytic Palmoplantar Keratoderma

Objective:To investigate the effect of keratin 9 gene on the pathogenesis of epidermolysis palmoplantar keratoderma by siRNA targeting RNA interference(RNAi)technique silenced HaCaT cell line KRT9R163W expression.Methods:l.we construct plasmid with KRT9R163W and transfect to HaCaT cells,and make overexpressed.2.Designed for the specific siRNA KRT9 R163W gene by liposome transfected HaCaT cells,and the establishment of a negative control group,positive control group and the control group.3.fluorescence quantitative polymerase chain reaction(qPCR)method to detect cell gene silencing effect.Results:KRT9 R163WsiRNA HaCaT cells 24h after transfection,RT-PCR showed the mRNA of K9-R163W and KRT9-R163W +siRNA-R163W relatively express 1.063±0.159,0.242±0.051;protein relatively express 1.1329±10.131,0.289±0.036;siRNA can siRNA can specificly inhibit mutant KRT9 expression(P<0.05with 95%confidence limits).The K9 gene was transfected by siRNA,the normal skeletal structure of the cells was changed into intricate reticular structure.Conclusion:Endogenous KRT9 is an important protein to maintain the normal skeletal structure of cells.SiRNA can specifically silence the expression of KRT9 R163W gene in HaCaT cells and provide a new theoretical basis for the study of pathogenesis of epidermolysis palmoplantar keratosis and target therapy.