Knockdown of H19 Enhances Differentiation Capacity to Epidermis of Parthenogenetic Embryonic Stem Cells

Yin, Yafei; Wang, H.; Liu, K.; Wang, F.; Ye, X.; Liu, M.; Xiang, Rong; Liu, N.; Liu, L.

doi:10.2174/1566524014666140724101035

Cited by 11 publications

(9 citation statements)

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“…In RWPE-1 cell line, H19 could confer stemness phenotype by these multiple pathways. In addition, knockdown of H19 in parthenogenetic embryonic stem cells (pES) promotes differentiation of pES to epidermis, showing that H19 RNA contributes to stem cell integrity [55].…”

Section: S-shipmentioning

confidence: 99%

Enrichment of Human Stem-Like Prostate Cells with s-SHIP Promoter Activity Uncovers a Role in Stemness for the Long Noncoding RNA H19

Roy

Vennin

Brocqueville

et al. 2015

Stem Cells and Development

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Understanding normal and cancer stem cells should provide insights into the origin of prostate cancer and their mechanisms of resistance to current treatment strategies. In this study, we isolated and characterized stem-like cells present in the immortalized human prostate cell line, RWPE-1. We used a reporter system with green fluorescent protein (GFP) driven by the promoter of s-SHIP (for stem-SH2-domain-containing 5¢-inositol phosphatase) whose stem cell-specific expression has been previously shown. We observed that s-SHIP-GFPexpressing RWPE-1 cells showed stem cell characteristics such as increased expression of stem cell surface markers (CD44, CD166, TROP2) and pluripotency transcription factors (Oct4, Sox2), and enhanced sphereforming capacity and resistance to arsenite-induced cell death. Concomitant increased expression of the long noncoding RNA H19 was observed, which prompted us to investigate a putative role in stemness for this oncofetal gene. Targeted suppression of H19 with siRNA decreased Oct4 and Sox2 gene expression and colonyforming potential in RWPE-1 cells. Conversely, overexpression of H19 significantly increased gene expression of these two transcription factors and the sphere-forming capacity of RWPE-1 cells. Analysis of H19 expression in various prostate and mammary human cell lines revealed similarities with Sox2 expression, suggesting that a functional relationship may exist between H19 and Sox2. Collectively, we provide the first evidence that s-SHIP-GFP promoter reporter offers a unique marker for the enrichment of human stem-like cell populations and highlight a role in stemness for the long noncoding RNA H19.

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Section: S-shipmentioning

confidence: 99%

Enrichment of Human Stem-Like Prostate Cells with s-SHIP Promoter Activity Uncovers a Role in Stemness for the Long Noncoding RNA H19

Roy

Vennin

Brocqueville

et al. 2015

Stem Cells and Development

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“…Recently, Yin et al . demonstrated that knockdown of H19 can improve mouse pESC differentiation potential in the lineages of ectoderm and mesoderm [ 50 ]. In further study, H19-Ifg2 and Dlk1-Dio3 imprinting region co-modification may have the greatest impact on improvement of the differentiation ability of pESCs.…”

Section: Discussionmentioning

confidence: 99%

Ascorbic acid improves pluripotency of human parthenogenetic embryonic stem cells through modifying imprinted gene expression in the Dlk1-Dio3 region

Gao

Zhao

et al. 2015

Stem Cell Res Ther

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IntroductionHuman parthenogenetic embryonic stem cells (hpESCs) are generated from artificially activated oocytes, however, the issue of whether hpESCs have equivalent differentiation ability to human fertilized embryonic stem cells remains controversial.MethodshpESCs were injected into male severe combined immunodeficiency (SCID) mice and the efficiency of teratoma formation was calculated. Then the gene expression and methylation modification were detected by real time-PCR and bisulfate methods.ResultsComparison of five hpESCs with different differentiation abilities revealed that levels of paternal genes in the Dlk1-Dio3 region on chromosome 14 in the hpESCs with high differentiation potential are enhanced, but strictly methylated and silenced in the hpESCs with lower differentiation potential. Treatment with ascorbic acid, rescued their ability to support teratoma formation and altered the expression profiles of paternally expressed genes in hpESCs that could not form teratoma easily. No differences in the expression of other imprinting genes were evident between hpESCs with higher and lower differentiation potential, except for those in the Dlk1-Dio3 region.ConclusionsThe Dlk1-Dio3 imprinting gene cluster distinguishes the differentiation ability of hpESCs. Moreover, modification by ascorbic acid may facilitate application of hpESCs to clinical settings in the future by enhancing their pluripotency.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0054-9) contains supplementary material, which is available to authorized users.

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“…Similarly, the results presented by Gabory et al confirmed that H19 played a negative role in cell proliferation [11]. Lately, it was reported that knockdown of H19 accelerated differentiation of parthenogenetic embryonic stem cells (p-ESCs) to cardiomyocytes [16, 17]. Nevertheless, little is known about the precise molecular mechanisms of H19 in cardiomyocyte differentiation.…”

Section: Resultsmentioning

confidence: 77%

Downregulation of long non-coding RNA H19 promotes P19CL6 cells proliferation and inhibits apoptosis during late-stage cardiac differentiation via miR-19b-modulated Sox6

Han

Cheng

et al. 2016

Cell Biosci

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BackgroundRegulating cardiac differentiation to maintain normal heart development and function is very important. At present, biological functions of H19 in cardiac differentiation is not completely clear.MethodsTo explore the functional effect of H19 during cardiac differentiation. Expression levels of early cardiac-specific markers Nkx-2.5 and GATA4, cardiac contractile protein genes α-MHC and MLC-2v were determined by qRT-PCR and western lot. The levels of lncRNA H19 and miR-19b were detected by qRT-PCR. We further predicted the binding sequence of H19 and miR-19b by online softwares starBase v2.0 and TargetScan. The biological functions of H19 and Sox6 were evaluated by CCK-8 kit, cell cycle and apoptosis assay and caspase-3 activity.ResultsThe expression levels of α-MHC, MLC-2v and H19 were upregulated, and miR-19b was downregulated significantly in mouse P19CL6 cells at the late stage of cardiac differentiation. Biological function analysis showed that knockdown of H19 promoted cell proliferation and inhibits cell apoptosis. H19 suppressed miR-19b expression and miR-19b targeted Sox6, which inhibited cell proliferation and promoted apoptosis in P19CL6 cells during late-stage cardiac differentiation. Importantly, Sox6 overexpression could reverse the positive effects of H19 knockdown on P19CL6 cells.ConclusionDownregulation of H19 promoted cell proliferation and inhibited cell apoptosis during late-stage cardiac differentiation by regulating the negative role of miR-19b in Sox6 expression, which suggested that the manipulation of H19 expression could serve as a potential strategy for heart disease.

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Knockdown of H19 Enhances Differentiation Capacity to Epidermis of Parthenogenetic Embryonic Stem Cells

Cited by 11 publications

References 0 publications

Enrichment of Human Stem-Like Prostate Cells with s-SHIP Promoter Activity Uncovers a Role in Stemness for the Long Noncoding RNA H19

Enrichment of Human Stem-Like Prostate Cells with s-SHIP Promoter Activity Uncovers a Role in Stemness for the Long Noncoding RNA H19

Ascorbic acid improves pluripotency of human parthenogenetic embryonic stem cells through modifying imprinted gene expression in the Dlk1-Dio3 region

Downregulation of long non-coding RNA H19 promotes P19CL6 cells proliferation and inhibits apoptosis during late-stage cardiac differentiation via miR-19b-modulated Sox6

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