Abstract
Human dental pulp stem cells (hDPSCs) have been recognized as a candidate cell source for tissue engineering. Long non-coding RNAs (lncRNAs) are differentially expressed in inflamed human dental pulp tissues. The present study is aimed at investigating the role of lncRNA H19 in the differentiation potential of hDPSCs. hDPSCs were successfully isolated and cultured, followed by conducting gain and loss-of-function experiments on lncRNA H19 and large tumor suppressor 1 (LATS1) to elucidate their respective biological functions in hDPSCs. lncRNA H19 was able to promote, whereas LATS1 was found to inhibit the differentiation, proliferation, and migration capabilities of hDPSCs. LATS1 was found to activate the Hippo-Yes-associated protein (YAP) signaling pathway by decreasing levels of YAP and Tafazzin (TAZ). The effects of lncRNA H19 on hDPSCs were achieved by repressing LATS1 through enhancer of zeste homolog 2-induced trimethylation of histone 3 at lysine 27. Finally, hDPSCs overexpressing lncRNA H19 and/or LATS1 were transplanted into nude mice. It was shown that lncRNA H19 inhibited LATS1 to promote the production of odontoblasts in vivo. Taken together, lncRNA H19 serves as a contributor to the differentiation potential of hDPSCs via the inhibition of LATS1, therefore highlighting novel therapeutic targets for dental pulp repair.
Highlights
Human dental pulp stem cells are primarily produced by pulp tissues of permanent and deciduous third molar teeth and primary incisors.[1] hDPSCs are plastic-adherent cells with fibroblastlike morphology[2] and possess a pronounced proliferation potential and differentiation capacity;[3] they possess clonogenic and proliferative abilities with differentiation potential into epithelial, neurogenic, adipogenic, osteogenic, and chondrogenic lineages.[4]
The results of quantitative real-time polymerase chain reaction (PCR), western blot analysis, and alkaline phosphatase (ALP) activity detection illustrated that overexpressing Long non-coding RNAs (lncRNAs) H19 had elevated the expression of odontoblast differentiation markers including ALP, dentin sialophosphoprotein (DSPP), osteocalcin (OCN), and dentin matrix protein 1 (DMP1) (Figures 2A, 2B, and S2A)
No significant differences were witnessed regarding the above-mentioned cellular interactions in hDPSCs without any treatment when compared with the transfection of negative control (NC) for the overexpressed plasmid and short hairpinRNA targeting H19, suggesting plasmid transfection had no impact on experimental data
Summary
Human dental pulp stem cells (hDPSCs) are primarily produced by pulp tissues of permanent and deciduous third molar teeth and primary incisors.[1] hDPSCs are plastic-adherent cells with fibroblastlike morphology[2] and possess a pronounced proliferation potential and differentiation capacity;[3] they possess clonogenic and proliferative abilities with differentiation potential into epithelial, neurogenic, adipogenic, osteogenic, and chondrogenic lineages.[4]. The detailed molecular mechanism underpinning this function still remains largely unknown
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