Project description:Human primary keratinocytes were depleted of MLL2 by siRNA and induced to differentiated for 2 days by addition of Calcium Primary normal human keratinocytes were transfected with MLL2 or scrambled control siRNA using RNAi max (Life Technologies). 24 hours post transfection medium was raised to 1.8mM calcium to induce differentiation. Cells were collected 48 hours later.
Project description:Human primary keratinocytes were depleted of GRHL3 by siRNA and induced to differentiated for 2 days by addition of Calcium Primary normal human keratinocytes were transfected with GRHL3 or scrambled control siRNA using RNAi max (Life Technologies). 24 hours post transfection medium was raised to 1.8mM to induce differentiation. Cells were collected 48 hours later.
Project description:Long non-coding RNAs (lncRNAs) have emerged as important regulators of biological processes and diseases, the function of the majority of epidermal lncRNAs in skin homeostasis and their potential role in psoriasis has remained unknown. We report the identification of lncRNAs with altered expression in psoriasis keratinocytes and identify LINC00958 is overexpressed in psoriasis epidermis. We used microarrays to detail the global programme of gene expression upon LINC00958 depletion in keratinocytes
Project description:The genetic expression profile of a Wnt signal agonist, BIO, was evaluated in human primary keratinocytes. Accelerating scaling up of primary keratinocytes benefits skin autografts for severely burned patients. Wnt signal, a conserved pathway controlling cell cycle and morphogenesis of embryo, has been postulated to promote the cell proliferation and tumorigenesis in adult. Here, the effects of Wnt signal on the growth of interfollicular keratinocytes were investigated. We demonstrated that recombinant Wnt3a significantly promoted the primary keratinocyte growth at a low cell density. A well-characterized GSK-3beta inhibitor, BIO, activated the Wnt signals and also enhanced the colony formation of keratinocytes dose-dependently. Gene expression profile of the BIO-treated keratinocytes revealed the linkage of the BIO with the cell mitosis and indicated that epithelial cell adhesion molecule (EpCAM), a Wnt target gene, was upregulated. Comparing to the EpCAM- keratinocytes, the EpCAM+ cells showed higher proliferation rate and efficacy of the colony formation. Especially, inhibiting the EpCAM expression by shRNA attenuated the proliferation effect of BIO and the growth advantage of the EpCAM+ keratinocytes. These evidences emphasize the positive role of canonical Wnt and EpCAM on the regulation of cell growth and self-renewal for human keratinocytes.
Project description:Sos proteins are involved in the maintaining of skin homeostasis and participate in the initiation/progression of chemically-induced skin cancer. However, the precise role of these proteins in specific skin cellular compartments remains undetermined. Here, we examined the role of Sos1 and Sos2 in the overall transcriptional profile of mouse primary keratinocytes in order to unveil the specific functions of these Ras activators in keratinocyte population. We used microarrays to detail the impact of Sos1/2 depletion (individual or combined) in the transcriptional signature of mouse primary keratinocytes