Gene expression and functional analysis of GJB2 mutated HaCaT cell lines [D66H]

Vohwinkel syndrome, VS (OMIM#124500), a rare autosomal dominant genetic disease, with less than 50 reported cases in the literature. Although clinical symptoms of VS are complex, which are caused by GJB2 mutation is more typical. To explore related differential genes and signaling pathways of Vohwinkel syndrome (VS) caused by mutations of GJB2. Human Gene Expression Array of two types of GJB2-VS mutated (G130V and D66H) HaCaT cell lines and identified distinct classes of up- and down- regulated genes during this process. Using GJB2 gene mutant p.Asp66His(D66H) primers as templates, the corresponding cDNA sequence of GJB2: NM_004004 (D66H) was amplified respectively, and then was ligated with GV308 vector. The recombinant plasmids D66H-GV308, and empty vector GV308 were identified and sequenced. Then GJB2-D66H-overexpression lentiviruses. Human Gene Expression Array of two types of GJB2-VS mutated (D66H) and NC HaCaT cell lines for RNA extraction and hybridization on Affymetrix microarrays. And combined Ingenuity Pathway Analysis, combined apoptosis experiment, mRNA expression of LOR, FLG, and differentially expressed genes.