분화유도제에 의한 감마 글로빈 유전자 프로모터에서의 DNA-단백질 상호작용

김덕인,박주인,김인후 大韓血液學會 1999 大韓血液學會誌 Vol.34 No.3

Role of Ras/ERK-dependent pathway in the erythroid differentiation of K562 cells

Kang, Chi-Dug,Do, In-Rok,Kim, Kwang-Woon,Ahn, Byung-Kwon,Kim, Sun-Hee,Chung, Byung-Seon,Jhun, Byung-Hak,Yoo, Mi-Ae 부산대학교 유전공학연구소 1999 분자생물학 연구보 Vol.15 No.-

The chronic myelogenous leukemic K562 cell line carrying Bcr-Abl tyrosine kinase is considered as pluripotent hematopoietic progenitor cells expressing markers for erythroid, granulocytic, monocytic, and megakaryocytid lineages. Here we investigated the signaling modulations required for induction of erythroid differentiation of K562 cells. When the K562 cells were treated with herbimycin A(an inhibitor of protein tyrosine kinase), ras antisense oligonucleotide, and PD98059 (a specific inhibitor of MEK), inhibition of ERK/MAPK activity and cell growth, and induction of erythroid differentiation were observed. The ras mutant, pZIPRas^61leu -transfected cells, K562-Ras^61leu have shown a markedly decreased cell proliferation rate with approximately 2-fold doubling time, compared with 2-fold doubling time, compared with the parental K562 cells, and about 60% of these cells have shown the phenotype of erythroid differentiation. In addition, herbimycin A inhibited the growth rate and increased the erythroid differentiation, but did not affect the elevated activity of ERK/MAPK in the K562-Ras^61leu cells. On the other hand, effects of PD98059 on the growth and differentiation of K562-Ras^61leu cells were biphasic. At low concentration of PD98059, which inhibited the elevated activity of ERK/MAPK to the level of parental cells, the growth rate increased and the erythroid differentiation decreased slightly, and at high concentration of PD98059, which inhibited the elevated activity of ERK/MAPK below that of the parental cells, the growth rate turned down and the erythroid differentiation was restored to the untreated control level. Taken together, these results suggest that an appropriate activity of ERK/MAPK is required to maintain the rapid growth and transformed phenotype of K562 cells.

The Up-Regulation of miR-199b-5p in Erythroid Differentiation Is Associated with GATA-1 and NF-E2

Yuxia Li,Hua Bai,Zhongzu Zhang,Weihua li,Lei Dong,Xueju Wei,Yanni Ma,Junwu Zhang,Jia Yu,Guotao Sun,Fang Wang 한국분자세포생물학회 2014 Molecules and cells Vol.37 No.3

MicroRNAs (miRNAs) represent a class of small non-co-ding regulatory RNAs that play important roles in normal hematopoiesis, including erythropoiesis. Although studies have identified several miRNAs that regulate erythroid commitment and differentiation, we do not understand the mechanism by which the crucial erythroid transcription factors, GATA-1and NF-E2 directly regulate and control differentiation via miRNA pathways. In this study, we identified miR-199b-5p as a key regulator of human erythropoiesis, and its expression was up-regulated during the erythroid differentiation of K562 cells. Furthermore, the increase of miR-199b-5p in erythroid cells occurred in a GATA-1- and NF-E2-dependent manner during erythrocyte maturation. Both GATA-1 and NF-E2 bound upstream of the miR-199b gene locus and activated its transcription. Forced expression of miRNA-199b-5p in K562 cells affected erythroid cell proliferation and maturation. Moreover, we identified c-Kit as a direct target of miR-199b-5p in erythroid cells. Taken together, our results establish a functional link among the erythroid transcription factors GATA-1/NF-E2, miR-199b-5p and c-Kit, and provide new insights into the coupling of transcription and post-transcription regulation in erythroid differentiation.

The Up-Regulation of miR-199b-5p in Erythroid Differentiation Is Associated with GATA-1 and NF-E2

Li, Yuxia,Bai, Hua,Zhang, Zhongzu,li, Weihua,Dong, Lei,Wei, Xueju,Ma, Yanni,Zhang, Junwu,Yu, Jia,Sun, Guotao,Wang, Fang Korean Society for Molecular and Cellular Biology 2014 Molecules and cells Vol.37 No.3

MicroRNAs (miRNAs) represent a class of small non-coding regulatory RNAs that play important roles in normal hematopoiesis, including erythropoiesis. Although studies have identified several miRNAs that regulate erythroid commitment and differentiation, we do not understand the mechanism by which the crucial erythroid transcription factors, GATA-1and NF-E2 directly regulate and control differentiation via miRNA pathways. In this study, we identified miR-199b-5p as a key regulator of human erythropoiesis, and its expression was up-regulated during the erythroid differentiation of K562 cells. Furthermore, the increase of miR-199b-5p in erythroid cells occurred in a GATA-1- and NF-E2-dependent manner during erythrocyte maturation. Both GATA-1 and NF-E2 bound upstream of the miR-199b gene locus and activated its transcription. Forced expression of miRNA-199b-5p in K562 cells affected erythroid cell proliferation and maturation. Moreover, we identified c-Kit as a direct target of miR-199b-5p in erythroid cells. Taken together, our results establish a functional link among the erythroid transcription factors GATA-1/NF-E2, miR-199b-5p and c-Kit, and provide new insights into the coupling of transcription and post-transcription regulation in erythroid differentiation.

Microarray Profiling of Genes Differentially Expressed during Erythroid Differentiation of Murine Erythroleukemia Cells

김철근,Hyen Seok Heo,Ju Hyun Kim,Young Jin Lee,Sung-Hyun Kim,조윤신 한국분자세포생물학회 2005 Molecules and cells Vol.20 No.1

Murine erythroleukemia (MEL) cells are widely used to study erythroid differentiation thanks to their ability to terminally differentiate in vitro in response to chemical induction. At the molecular level, not much is known of their terminal differentiation apart from activation of adult-type globin gene expression. We examined changes in gene expression during the terminal differentiation of these cells using microarray-based technology. We identified 180 genes whose expression changed significantly during differentiation. The microarray data were analyzed by hierarchical and k-means clustering and confirmed by semi-quantitative RT-PCR. We identified several genes including H1f0, Bnip3, Mgl2, ST7L, and Cbll1 that could be useful markers for erythropoiesis. These genetic markers should be a valuable resource both as potential regulators in functional studies of erythroid differentiation, and as straightforward cell type markers.

Negative Regulation of Erythroid Differentiation via the CBX8-TRIM28 Axis

Kim, Hyun Jeong,Park, Jin Woo,Kang, Joo-Young,Seo, Sang-Beom Korean Society for Molecular and Cellular Biology 2021 Molecules and cells Vol.44 No.7

Although the mechanism of chronic myeloid leukemia (CML) initiation through BCR/ABL oncogene has been well characterized, CML cell differentiation into erythroid lineage cells remains poorly understood. Using CRISPR-Cas9 screening, we identify Chromobox 8 (CBX8) as a negative regulator of K562 cell differentiation into erythrocytes. CBX8 is degraded via proteasomal pathway during K562 cell differentiation, which activates the expression of erythroid differentiation-related genes that are repressed by CBX8 in the complex of PRC1. During the differentiation process, the serine/threonine-protein kinase PIM1 phosphorylates serine 196 on CBX8, which contributes to CBX8 reduction. When CD235A expression levels are analyzed, the result reveals that the knockdown of PIM1 inhibits K562 cell differentiation. We also identify TRIM28 as another interaction partner of CBX8 by proteomic analysis. Intriguingly, TRIM28 maintains protein stability of CBX8 and TRIM28 loss significantly induces proteasomal degradation of CBX8, resulting in an acceleration of erythroid differentiation. Here, we demonstrate the involvement of the CBX8-TRIM28 axis during CML cell differentiation, suggesting that CBX8 and TRIM28 are promising novel targets for CML research.

Disialoganglioside GD3 synthase expression recruits membrane transglutaminase 2 during erythroid differentiation of the human chronic myelogenous leukemia K562 cells

Kang, Sung-Koo,Kim, Yong-Sam,Kong, Yun-Jeong,Song, Kwon-Ho,Chang, Young-Chae,Park, Young-Guk,Ko, Jeong-Heon,Lee, Young-Choon,Kim, Cheorl-Ho WILEY-VCH Verlag 2008 Proteomics Vol.8 No.16

<P>By employing proteomics analysis tool, we examined the effects of GD3 synthase expression on the differentiation properties of chronic myelogenous leukemia (CML)-derived leukemia cells K562. Forced expression of GD3 synthase induced erythroid differentiation as determined by an increase in glycophorin A expression and synthesis of hemoglobins. The proteomic analysis revealed that 15 proteins were increased by GD3 synthase. In contrast, we observed three protein gel spots decreased in contents in the cell membranes of GD3 synthase-transfected K562 cells. Among the increased proteins, membrane transglutaminase 2 (TG2) was specifically increased in the cell membrane of GD3 synthase-transfected K562 cells. Then, we generated the GD3 synthase-transfected cells in the K562 cells. Interestingly, the TG2 level was increased in GD3 synthase-transfected cells compared with vector- and plasma membrane-associated ganglioside sialidase (Neu3)-transfected cells. In addition, its ability to be photoaffinity-labeled with [α-<SUP>32</SUP>P]GTP was also increased in the GD3 synthase- and TG2-transfected cells. Moreover, small interfering RNA (siRNA) analysis for the GD3 synthase showed the decrease or abolishment of the membrane TG2. Finally, GD3 synthase-transfected cells accelerated the erythroid differentiation. Therefore, we propose that the recruitment of TG2 into membranes by GD3 might play an important role in the erythroid differentiation in K562 cells.</P>

Activation of Nrf2 by sulfuretin stimulates chondrocyte differentiation and increases bone lengths in zebrafish

Seo-Hyuk Chang,Hoi-Khoanh Giong,Da-Young Kim,Suji Kim,Seungjun Oh,Ui Jeong Yun,이정수,Kye Won Park 생화학분자생물학회 2023 BMB Reports Vol.56 No.9

Elongation of most bones occur at the growth plate throughendochondral ossification in postnatal mammals. The maturationof chondrocyte is a crucial factor in longitudinal bone growth,which is regulated by a complex network of paracrine and endocrinesignaling pathways. Here, we show that a phytochemicalsulfuretin can stimulate hypertrophic chondrocyte differentiationin vitro and in vivo. We found that sulfuretin stabilizednuclear factor (erythroid-derived 2)-like 2 (Nrf2), stimulatedits transcriptional activity, and induced expression of itstarget genes. Sulfuretin treatment resulted in an increase inbody length of zebrafish larvae and induced the expression ofchondrocyte markers. Consistently, a clinically available Nrf2activator, dimethyl fumarate (DMF), induced the expression ofhypertrophic chondrocyte markers and increased the body lengthof zebrafish. Importantly, we found that chondrocyte gene expressionin cell culture and skeletal growth in zebrafish stimulatedby sulfuretin were significantly abrogated by Nrf2 depletion,suggesting that such stimulatory effects of sulfuretin weredependent on Nrf2, at least in part. Taken together, these datashow that sulfuretin has a potential use as supporting ingredientsfor enhancing bone growth.

홍삼 비사포닌 분획의 단핵세포 분화와 염증반응에 대한 억제효과

강보빈(Bobin Kang),김채영(Chae Young Kim),황지수(Jisu Hwang),최현선(Hyeon-Son Choi) 한국식품과학회 2019 한국식품과학회지 Vol.51 No.1

본 연구에서는 홍삼 비사포닌 분획(NSF)의 항 염증 효과를 마우스 대식세포와 인간유래 단핵세포에서 확인하였다. NSF는 마우스 대식세포에서 LPS로 유도된 NO, iNOS 그리고 COX-2의 양 뿐만 아니라 IL-6, TNF-α, MCP-1과 같은 염증성 싸이토카인의 생성량을 유의적으로 감소시켰다. 인간 유래 단핵세포에서는 PMA에 의해 유도되는 대식세포로의 분화를 효과적으로 억제하면서 분화인자인 CD11β와 CD36의 발현을 유의적으로 감소시켰다. 마우스 대식세포에서와 마찬가지로 염증성 싸이토카인들의 생성량 또한 감소하였는데, 이러한 NSF의 항 염증 효과는 두 전사인자의 조절작용에 의한 것으로 사료된다. 즉 NSF는 NF-κB의 핵으로 이동을 감소시킴으로써 전사활성을 억제하여 염증성 싸이토카인들의 발현을 저해하고 이와 반대로 Nrf2의 발현과 핵으로의 이동을 증가시켜 항산화 효소이면서 항 염증 작용을 나타내는 HO-1의 발현을 촉진하는 것으로 관찰되었다. 따라서 NSF는 NF-κB와 Nrf2의 두 가지 신호전달체계를 조절함으로써 항 염증 작용을 나타냈으며 이를 홍삼 NSF의 항 염증 기작으로 보고하는 바이다. The aim of this study was to investigate the effects of red ginseng-derived non-saponin fraction (NSF) on inflammatory responses and monocyte-to-macrophage differentiation in RAW264.7 and THP-1. NSF effectively inhibited inflammatory responses by downregulating nitric oxide (NO) production and protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). NSF (2000 μg/mL) decreased the levels of NO, iNOS, and COX-2 by 33, 83, and 64%, respectively. NSF inhibited the differentiation of monocyte-to-macrophage by decreasing cell adherence along with downregulation of the cluster of differentiation molecule 11β (CD11β) and CD36. In addition, pro-inflammatory cytokines, such as tumor necrosis factor-alpha, interleukin 6, and monocyte chemoattractant protein 1 (MCP-1), were significantly reduced with NSF treatment. The NSF-mediated inhibition of inflammatory responses was due to the regulation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 (Nrf2). NSF effectively suppressed the translocation of NF-κB into the nucleus, while nuclear Nrf2 and its target protein, heme oxygenase-1, levels were significantly increased.

P145 Erythroid differentiation regulator 1 expression in various types of alopecias

( Yu Ri Woo ),( Jong Sic Kim ),( Ji Hong Lim ),( Sae Won Hwang ),( Miri Kim ),( Hyun Jeong Park ) 대한피부과학회 2016 대한피부과학회 학술발표대회집 Vol.68 No.2

<div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div> Background: Erythroid differentiation regulator 1 (Erdr1) is known to involved in the inflammatory process via regulating immune system in various cutaneous disorders, psoriasis, rosacea. However, the role of Erdr1 in alopecia has not been studied. Objectives: The aim of this study is to investigate the expression patterns of Erdr1 in various types of alopecias. Methods: In all, twenty patients with alopecia were included. Skin samples from nine patients with alopecia areata, eight patients with scarring alopecia, three patients with traumatic alopecia, and five control subjects were retrieved for evaluation of Erdr1 expression. Results: Downregulation of Erdr1 was observed in alopecia than control groups. The expression density of Erdr1 was weaker in hair follicle than epidermis. In particular, a significant decrease in expression density of Erdr1 was observed in 44% of patients with alopecia areata, who were recalcitrant to the treatment. Conclusion: The expression of Erdr1 was downregulated in alopecia, which suggest that Erdr1 may be involved in the pathogenesis. Especially for alopecia areata, the expression of Erdr1 could be used as a prognostic factor for disease progression and treatment outcome.