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Authors:Zou Yuxi, Sun Jing, Sun Yu, Suo Xi, Zhou Wenkai, Yang Jianguang, Liu Yan
Keywords:Lin28;mTOR;human dental pulp stem cells;proliferation;osteogenic differentiation
DOI:10.19405/j.cnki.issn1000-1492.2023.09.012
〔Abstract〕 Objective To investigate the effects of Lin28 overexpression on the proliferation and osteogenic differentiation of human dental pulp stem cells(hDPSCs) through mTOR signaling pathway. Methods After transfecting lentiviral vectors of Lin28 gene in hDPSCs, the relative expression of Lin28 was detected by Real-time PCR. CCK-8 assay was applied to detect the effect on cell proliferation. qRT-PCR was used to research the expression levels of alkaline phosphatase(ALP), osteopontin(OPN) and osteocalcin(OCN). Western blot assay was processed to investigate the effects on the relative expression levels of ALP and OPN proteins. Alizarin red staining was utilized to detect the mineralized nodules. Results Compared with the control group, the cell proliferation of transfection group was promoted(P<0.05); The mRNA and protein expression levels of ALP, OPN and OCN in transfection group were significantly lower than those in control group(P<0.05), the expression level of ALP apparently decreased after the addition of mTOR inhibitor rapamycin(P<0.05); Alizarin red staining showed that the size and number of mineralized nodules formed in transfection group were markedly declined compared with empty carrier group(P<0.05). Conclusion Overexpression of Lin28 can inhibit the osteogenic differentiation of hDPSCs through suppress mTOR signaling pathway.