LIN28A,an RNA-binding protein,plays an important role in porcine induced pluripotent stem cells(piPSCs).However,the molecular mechanism underlying the function of LIN28A in the maintenance of pluripotency in piPSCs re...LIN28A,an RNA-binding protein,plays an important role in porcine induced pluripotent stem cells(piPSCs).However,the molecular mechanism underlying the function of LIN28A in the maintenance of pluripotency in piPSCs remains unclear.Here,we explored the function of LIN28A in piPSCs based on its overexpression and knockdown.We performed total RNA sequencing(RNA-seq)of piPSCs and detected the expression levels of relevant genes by quantitative real-time polymerase chain reaction(qRT-PCR),western blot analysis,and immunofluorescence staining.Results indicated that piPSC proliferation ability decreased following LIN28A knockdown.Furthermore,when LIN28A expression in the shLIN28A2 group was lower(by 20%)than that in the negative control knockdown group(shNC),the pluripotency of piPSCs disappeared and they differentiated into neuroectoderm cells.Results also showed that LIN28A overexpression inhibited the expression of DUSP(dual-specificity phosphatases)family phosphatases and activated the mitogen-activated protein kinase(MAPK)signaling pathway.Thus,LIN28A appears to activate the MAPK signaling pathway to maintain the pluripotency and proliferation ability of piPSCs.Our study provides a new resource for exploring the functions of LIN28A in piPSCs.展开更多
As a transcription factor of the Pit-Oct-Unc(POU)domain family,octamer-binding transcription factor 6(OCT6)participates in various aspects of stem cell development and differentiation.At present,however,its role in po...As a transcription factor of the Pit-Oct-Unc(POU)domain family,octamer-binding transcription factor 6(OCT6)participates in various aspects of stem cell development and differentiation.At present,however,its role in porcine-induced pluripotent stem cells(piPSCs)remains unclear.Here,we explored the function of OCT6 in piPSCs.We found that piPSCs overexpressing OCT6 maintained colony morphology and pluripotency under differentiation conditions,with a similar gene expression pattern to that of non-differentiated piPSCs.Functional analysis revealed that OCT6 attenuated the adverse effects of extracellular signal-regulated kinase(ERK)signaling pathway inhibition on piPSC pluripotency by activating phosphatidylinositol 3-kinase-protein kinase B(PI3K-AKT)signaling activity.Our research sheds new light on the mechanism by which OCT6 promotes PSC maintenance.展开更多
基金This work was supported by the National Key Research,Development Program of China-Stem Cell and Translational Research(2016YFA0100200)National Natural Science Foundation of China(32072806,31572399,61772431,62072377)+1 种基金Program of Shaanxi Province Science and Technology Innovation Team(2019TD-036)Fundamental Research Funds for the Central Universities,Northwest A&F University(Z1090219146,Z102022004)。
文摘LIN28A,an RNA-binding protein,plays an important role in porcine induced pluripotent stem cells(piPSCs).However,the molecular mechanism underlying the function of LIN28A in the maintenance of pluripotency in piPSCs remains unclear.Here,we explored the function of LIN28A in piPSCs based on its overexpression and knockdown.We performed total RNA sequencing(RNA-seq)of piPSCs and detected the expression levels of relevant genes by quantitative real-time polymerase chain reaction(qRT-PCR),western blot analysis,and immunofluorescence staining.Results indicated that piPSC proliferation ability decreased following LIN28A knockdown.Furthermore,when LIN28A expression in the shLIN28A2 group was lower(by 20%)than that in the negative control knockdown group(shNC),the pluripotency of piPSCs disappeared and they differentiated into neuroectoderm cells.Results also showed that LIN28A overexpression inhibited the expression of DUSP(dual-specificity phosphatases)family phosphatases and activated the mitogen-activated protein kinase(MAPK)signaling pathway.Thus,LIN28A appears to activate the MAPK signaling pathway to maintain the pluripotency and proliferation ability of piPSCs.Our study provides a new resource for exploring the functions of LIN28A in piPSCs.
基金supported by the National Natural Science Foundation of China(32072806)Shaanxi Province Science and Technology Innovation Team Project(2019TD-036)Shaanxi Province Science and Technology Project(2022NY-044)。
文摘As a transcription factor of the Pit-Oct-Unc(POU)domain family,octamer-binding transcription factor 6(OCT6)participates in various aspects of stem cell development and differentiation.At present,however,its role in porcine-induced pluripotent stem cells(piPSCs)remains unclear.Here,we explored the function of OCT6 in piPSCs.We found that piPSCs overexpressing OCT6 maintained colony morphology and pluripotency under differentiation conditions,with a similar gene expression pattern to that of non-differentiated piPSCs.Functional analysis revealed that OCT6 attenuated the adverse effects of extracellular signal-regulated kinase(ERK)signaling pathway inhibition on piPSC pluripotency by activating phosphatidylinositol 3-kinase-protein kinase B(PI3K-AKT)signaling activity.Our research sheds new light on the mechanism by which OCT6 promotes PSC maintenance.
