Sox6 overexpression causes cellular aggregation and the neuronal differentiation of P19 embryonic carcinoma cells in the absence of retinoic acid

Hamada-Kanazawa, Michiko; Ishikawa, Kyoko; Nomoto, Kaori; Uozumi, Takako; Kawai, Yuichi; Narahara, Masanori; Miyake, Masanobu

doi:10.1016/s0014-5793(04)00086-9

Cited by 34 publications

(38 citation statements)

References 26 publications

(42 reference statements)

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“…1D), mRNA transcripts of N-cadherin (a neuron-specific cell adhesion molecule) and ␤III-tubulin (Fig. 1C) gradually increased with differentiation of the P19 cells, as observed previously (20,26,54). These two neuronal markers were also used for either Northern or Western blots ( Fig.…”

Section: Induction Of the Opioid Gene During P19 Cell Differentiationsupporting

confidence: 73%

Evidence of Endogenous Mu Opioid Receptor Regulation by Epigenetic Control of the Promoters

Hwang

Song

Kim

et al. 2007

Molecular and Cellular Biology

145

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The pharmacological effect of morphine as a painkiller is mediated mainly via the mu opioid receptor (MOR) and is dependent on the number of MORs in the cell surface membrane. While several studies have reported that the MOR gene is regulated by various cis-and trans-acting factors, many questions remain unanswered regarding in vivo regulation. The present study shows that epigenetic silencing and activation of the MOR gene are achieved through coordinated regulation at both the histone and DNA levels. In P19 mouse embryonal carcinoma cells, expression of the MOR was greatly increased after neuronal differentiation. MOR expression could also be induced by a demethylating agent (5-aza-2-deoxycytidine) or histone deacetylase inhibitors in the P19 cells, suggesting involvement of DNA methylation and histone deacetylation for MOR gene silencing. Analysis of CpG DNA methylation revealed that the proximal promoter region was unmethylated in differentiated cells compared to its hypermethylation in undifferentiated cells. In contrast, the methylation of other regions was not changed in either cell type. Similar methylation patterns were observed in the mouse brain. In vitro methylation of the MOR promoters suppressed promoter activity in the reporter assay. Upon differentiation, the in vivo interaction of MeCP2 was reduced in the MOR promoter region, coincident with histone modifications that are relevant to active transcription. When MeCP2 was disrupted using MeCP2 small interfering RNA, the endogenous MOR gene was increased. These data suggest that DNA methylation is closely linked to the MeCP2-mediated chromatin structure of the MOR gene. Here, we propose that an epigenetic mechanism consisting of DNA methylation and chromatin modification underlies the cell stage-specific mechanism of MOR gene expression.Opioids exert their pharmacological and physiological effects through binding to their endogenous receptors. Three types of opioid receptors, mu (), delta (␦), and kappa (), all belonging to the G-protein-coupled receptor superfamily, have been cloned. Upon agonist binding, these receptors couple to G proteins and affect several signal transduction pathways thought to mediate a broad range of functions and pharmacological effects of endogenous and exogenous opioids (51). Previous studies suggested that the opioid receptor (MOR) plays a key role in mediating the major clinical effects of analgesics, such as morphine, as well as the development of tolerance and physical dependence upon prolonged administration (39). MOR is mainly expressed in the central nervous system, with densities varying greatly in different regions, which can display different functional roles (55). During mouse embryonic development, the MOR message was specifically observed as early as embryonic day 8.5 (E8.5) using the reverse transcription (RT)-PCR method (44). In contrast, MOR transcripts were detected only beginning at E12 using the radioligand binding method (70) and at E10.5 by in situ hybridization (85). Transcript levels gradually incr...

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Section: Induction Of the Opioid Gene During P19 Cell Differentiationsupporting

confidence: 73%

Evidence of Endogenous Mu Opioid Receptor Regulation by Epigenetic Control of the Promoters

Hwang

Song

Kim

et al. 2007

Molecular and Cellular Biology

145

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“…Actually, this conclusion has been proved by some experiments. Altering the expression of preneural genes can promote P19 cell differentiating without the RA treatment (such as Sox-6 and Wnt-1 [24,25]) or without the cell aggregation (such as FGF8 [30]). Considering that cell aggregation is also essential for differentiation of P19 cells into muscle cells when treated with DMSO, it is possible that aggregation can promote the pluripotent undifferentiated P19 cells to become more differentiated cells, but not enough to neural cell linage [29].…”

Section: Discussionmentioning

confidence: 99%

“…But under some circumstances, gene modification can promote P19 cells to differentiate into neurons in the absence of RA, such as Sox-6 and WNT-1 overexpression [24,25]. After P19/23B10 cells have aggregated for 4 days, without RA treatment, the disassociated P19/23B10 cells began to extrude neurite-like structure on D2 (Fig.…”

Section: Choosing the Rnai Target Sequence For Adam23mentioning

confidence: 99%

ADAM23 Plays Multiple Roles in Neuronal Differentiation of P19 Embryonal Carcinoma cells

et al. 2007

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ADAM23, belonging to ADAM (A Disintegrin And Metalloprotease) protein family, is mainly expressed in brain. P19 cells could differentiate into neuroectodermal cell lineage after cell aggregates have been induced by retinoic acid (RA). In this report, we show that the post-transcriptional and post-translational processes of ADAM23 are regulated during the differentiation of P19 cells. In P19-derived neurons, ADAM23 is polarized distributed in the proximal part. To explore the possible roles of ADAM23 during P19 cell neuronal differentiation, ADAM23-RNAi P19 cell lines were established. These transfected cells could differentiate into neurofilament-expression neurons in the absence of RA, whereas wild-type P19 cell can not. These results suggest ADAM23 may play roles in both early and later stage of neuronal differentiation.

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“…[1][2][3] In particular, Sox6, originally isolated from adult mouse testis, 4 is required for the development of the central nervous system, [5][6][7] for chondrogenesis, 8 and for cardiac and skeletal muscle formation. 9,10 Recently, Sox6 has been demonstrated to be crucial for definitive erythropoiesis, [11][12][13][14][15] a process in which committed progenitors progressively differentiate into burst-forming-unit erythroid cells and colonyforming-unit (CFU) erythroid cells, which in turn give rise to proerythroblasts and erythroblasts and finally to mature, enucleated red blood cells.…”

Section: Introductionmentioning

confidence: 99%

“…The Sox6 recombinant protein lacks the 49 C-terminal amino acids; this shorter molecule fully retains Sox6 biologic properties. 6 The Sox6FLAG cassette (EcoRI-KpnI-blunted sites) was then cloned immediately upstream of the internal ribosomal entry site (IRES)-emerald green fluorescent protein (GFP) cassette (blunted BamHI site) of the pHR-SIN-BX-IR/EMW vector (derived from pHR-SIN-CSGW lentiviral vector 21 ). The TWEEN vector containing human SOCS3 cDNA was kindly provided by Dr M. G. Francipane (University of Palermo, Italy).…”

mentioning

confidence: 99%

Sox6 enhances erythroid differentiation in human erythroid progenitors

Cantù

Ierardi

Alborelli

et al. 2011

Blood

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Sox6 belongs to the Sry (sex-determining region Y)-related high-mobility-groupbox family of transcription factors, which control cell-fate specification of many cell types. Here, we explored the role of Sox6 in human erythropoiesis by its overexpression both in the erythroleukemic K562 cell line and in primary erythroid cultures from human cord blood CD34 ؉ cells. Sox6 induced significant erythroid differentiation in both models. K562 cells underwent hemoglobinization and, despite their leukemic origin, died within 9 days after transduction; primary erythroid cultures accelerated their kinetics of erythroid maturation and increased the number of cells that reached the final enucleation step. Searching for direct Sox6 targets, we found SOCS3 (suppressor of cytokine signaling-3), a known mediator of cytokine response. Sox6 was bound in vitro and in vivo to an evolutionarily conserved regulatory SOCS3 element, which induced transcriptional activation. SOCS3 overexpression in K562 cells and in primary erythroid cells recapitulated the growth inhibition induced by Sox6, which demonstrates that SOCS3 is a relevant Sox6 effector. (Blood. 2011;117(13):3669-3679) IntroductionSox proteins are important transcriptional regulators of different developmental processes in which they control the specification and differentiation of many cell types. [1][2][3] In particular, Sox6, originally isolated from adult mouse testis, 4 is required for the development of the central nervous system, 5-7 for chondrogenesis, 8 and for cardiac and skeletal muscle formation. 9,10 Recently, Sox6 has been demonstrated to be crucial for definitive erythropoiesis, 11-15 a process in which committed progenitors progressively differentiate into burst-forming-unit erythroid cells and colonyforming-unit (CFU) erythroid cells, which in turn give rise to proerythroblasts and erythroblasts and finally to mature, enucleated red blood cells. These differentiation stages are accompanied by profound maturational changes: Within few cell divisions, in parallel with the accumulation of erythroid-specific markers (membrane proteins, enzymes required for the heme biosynthesis pathway, and globins), cells undergo chromatin condensation and enucleate. 16,17 This complex spectrum of maturational steps is controlled at the molecular level by the integration of extrinsic (growth factors; oxygen and iron availability) and intrinsic (growth factor receptors, signaling mediators, transcription factors) signals.Several transcription factors are essential for erythroid commitment and for differential globin gene expression during development; their absence is associated with a wide spectrum of phenotypes ranging from mild perturbation to death because of a complete failure of erythropoiesis. 18,19 Among them, Sox6 recently has been shown to stimulate erythroid cell survival, proliferation, and terminal maturation during definitive murine erythropoiesis. 11,12 Sox6-null mouse fetuses and pups are anemic and have defective red blood cells. Recently, Sox6 has been implicated...

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Sox6 overexpression causes cellular aggregation and the neuronal differentiation of P19 embryonic carcinoma cells in the absence of retinoic acid

Cited by 34 publications

References 26 publications

Evidence of Endogenous Mu Opioid Receptor Regulation by Epigenetic Control of the Promoters

Evidence of Endogenous Mu Opioid Receptor Regulation by Epigenetic Control of the Promoters

ADAM23 Plays Multiple Roles in Neuronal Differentiation of P19 Embryonal Carcinoma cells

Sox6 enhances erythroid differentiation in human erythroid progenitors

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