RNAseq of TGF-β receptor type I kinase-dependent genes in oral fibroblast exposed to milk

Abstract

Background Milk is a rich source of natural growth factors that may support oral tissue homeostasis and wound healing. We had shown earlier that blocking TGF-β receptor type I kinase with the inhibitor SB431542 abolished the expression of IL11 and other genes in human gingival fibroblasts exposed to the aqueous fraction of milk. Our aim was to identify the entire signature of TGF-β receptor type I kinase-dependent genes regulated by the aqueous fraction of human milk. Result RNAseq revealed 99 genes being strongly regulated by milk requiring activation of the SB431542-dependent TGF-β receptor type I kinase. Among the SB431542-dependent genes is IL11 but also cadherins, claudins, collagens, potassium channels, keratins, solute carrier family proteins, transcription factors, transmembrane proteins, tumor necrosis factor ligand superfamily members, and tetraspanin family members. When focusing on our candidate gene, we could identify D609 to suppress IL11 expression, independent of phospholipase C, sphinosine-1 phosphate synthesis, and Smad-3 phosphorylation and its nuclear translocation. In contrast, genistein and blocking phosphoinositide 3-kinases by wortmannin and LY294002 increased the milk-induced IL11 expression in gingival fibroblasts. Conclusion Taken together, our data revealed TGF-β receptor type I kinase signaling to cause major changes of the genetic signature of gingival fibroblasts exposed to aqueous fraction of human milk.

PRF Lysates Enhance the Proliferation and Migration of Oral Squamous Carcinoma Cell Lines

“…We have previously implemented our established oral squamous carcinoma cell line HSC2 as a bioassay for testing the responsiveness to lysates prepared from PRF membranes, however, in the context of inflammation research [27,28]. Today's attempts to assess the response of HSC2 and other cell types to PRF lysates are based on RNA sequencing, an omics technology that provides insight into how the genetic signature of cells changes [29]. Similar to the understanding of the response of gingival fibroblasts to milk, it is now time to identify how HSC2 cells respond to PRF lysates based on RNAseq, followed by an analysis of genes regulating the cell cycle such as cyclin D1, Ki67, and PCNA [30].…”