1.Expression of histone demethylase lysine specific demethylase 1 in acute leukemia and its clinical significance.
Xiu-Mei LIN ; Wen-Ting ZHONG ; Chun-Li WANG ; Shun-Qing WANG
Journal of Experimental Hematology 2011;19(6):1348-1352
The aim of this study was to investigate the expression of histone demethylase lysine specific demethylase1 (LSD1) in patients with acute leukemia (AL) and its clinical significance. LSD1 protein expression level was detected by semi-quantitative Western blot in HL-60 and SHI-1 leukemia cell line, in bone marrow mononuclear cells of acute AL patients with different condition [new diagnosis, complete remission (CR) and relapse] and in patients with non malignant hematopathy (control). Clinical data of AL patient followed up was collected. The relationship of LSD1 expression level with clinical prognosis was analyzed. The results showed that in HL-60 and SHI-1 leukemia cell line, LSD1 expression was strong positive, relative amount (LSD1/β-actin gray level rate) was 4.647 ± 3.840 and 1.628 ± 0.185 (n = 4) respectively. In 72 AL patients, LSD1 expression levels were quite different. LSD1 positive rate was 56.9% (41/72), average relative amount was 1.053 ± 1.976. In 17 controls, LSD1 positive rate was 0%, relative amount was 0.004 ± 0.012. The LSD1 positive rate in newly diagnosed AML or ALL group (90.4%, 77.8%) and refractory/relapse AML or ALL group (100%, 100%) was higher than that in AML or ALL CR group (4.7%, 0%) (p = 0.000), relative amount of LSD1 showed no statistically difference between newly diagnosed AML and ALL groups (1.177 ± 1.646, 1.275 ± 1.845) or refractory/relapse group (2.050 ± 2.470, 4.107 ± 3.676) and CR group (0.029 ± 0.033, 0.019 ± 0.024) (p > 0.05). In all AL patients, LSD1 positive rate in newly diagnosed (84.6%) and refractory/relapse groups (100%) was higher than that in CR group (3.8%). LSD1 relative amount in newly diagnosed group (1.274 ± 1.760), refractory/relapse group (3.359 ± 3.319) and CR group (0.027 ± 0.031) was higher than that in control group (p < 0.01), and in refractory/relapse group was higher than that in newly diagnosed group and CR group (p < 0.01), in newly diagnosed group was higher than that in CR group (p < 0.01). It is concluded that overexpression of LSD1 is correlated with refractory or relapse in AL. LSD1 expression level can reflect disease status of AL patients and may be a predictive biomarker for unfavourable prognosis of AL.
HL-60 Cells
;
Histone Demethylases
;
metabolism
;
Humans
;
Leukemia
;
metabolism
;
Leukemia, Myeloid, Acute
;
metabolism
;
Recurrence
2.Research Advances in Lysine-specific Demethylase 6A and Its Application in Treating Leukemia.
Jie SHEN ; Zhi Hui ZENG ; Lei YANG ; Mao Jun ZENG ; Zhe Shen OUYANG ; Ming Yi ZHAO ; Ming Hua YANG
Acta Academiae Medicinae Sinicae 2019;41(4):548-555
Leukemia is a disease featured by the malignant proliferation of hematopoietic stem cells or progenitor cells in the blood system.While chemotherapy remains its mainstream treatment,disease relapse and drug resistance are still challenging problems.As one of the epigenetic mechanisms,histone methylation is involved in cell proliferation,differentiation,and apoptosis by regulating gene transcription.Recent studies have found that the histone demethylase lysine-specific demethylase 6A(KDM6A),also known as ubiquitously transcribed tetratricopeptide repeat on chromosome X(UTX),is closely related to the occurrence of a variety of tumors,especially leukemia.KDM6A activates gene expression by demethylating H3K27me3 to H3K27me2 or H3K27me1.Besides,KDM6A can regulate the activation of the target gene transcription through its non-demethylase functions.It can serve as the subunit of complex of proteins associated with Set1,thus getting involved in the regulation of H3K4me1.It can be combined with yeast mating type conversion/sucrose unfermented complex family to promote the formation of an open chromatin conformation.Finally,it can promote the production of H3K27ac.This article reviews the recent studies on the structure and biological activity of histone demethylase KDM6A(UTX)and its role in treating leukemia,thus providing a new research direction for targeted treatment of leukemia.
Epigenesis, Genetic
;
Histone Demethylases
;
metabolism
;
Histones
;
Humans
;
Leukemia
;
enzymology
;
therapy
;
Lysine
;
Nuclear Proteins
;
metabolism
3.Promising natural lysine specific demethylase 1 inhibitors for cancer treatment: advances and outlooks.
Zhong-Rui LI ; Meng-Zhen GU ; Xiao XU ; Jing-Han ZHANG ; Hai-Li ZHANG ; Chao HAN
Chinese Journal of Natural Medicines (English Ed.) 2022;20(4):241-257
Lysine specific demethylase 1 (LSD1), a transcriptional corepressor or coactivator that serves as a demethylase of histone 3 lysine 4 and 9, has become a potential therapeutic target for cancer therapy. LSD1 mediates many cellular signaling pathways and regulates cancer cell proliferation, invasion, migration, and differentiation. Recent research has focused on the exploration of its pharmacological inhibitors. Natural products are a major source of compounds with abundant scaffold diversity and structural complexity, which have made a major contribution to drug discovery, particularly anticancer agents. In this review, we briefly highlight recent advances in natural LSD1 inhibitors over the past decade. We present a comprehensive review on their discovery and identification process, natural plant sources, chemical structures, anticancer effects, and structure-activity relationships, and finally provide our perspective on the development of novel natural LSD1 inhibitors for cancer therapy.
Antineoplastic Agents/therapeutic use*
;
Enzyme Inhibitors/therapeutic use*
;
Histone Demethylases/metabolism*
;
Humans
;
Lysine/therapeutic use*
;
Neoplasms/drug therapy*
4.ZNF750 facilitates carcinogenesis via promoting the expression of long non-coding RNA CYTOR and influences pharmacotherapy response in colon adenocarcinoma.
Lu XIA ; Hexin LIN ; Yanming ZHOU ; Jiabian LIAN
Journal of Zhejiang University. Science. B 2022;23(7):587-596
The epidermal cell differentiation regulator zinc finger protein 750 (ZNF750) is a transcription factor containing the Cys2His2 (C2H2) domain, the zinc finger structure of which is located at the N-terminal 25-46 amino acids of ZNF750. It can promote the expression of differentiation-related factors while inhibiting the expression of progenitor cell-related genes. ZNF750 is directly regulated by p63 (encoded by the TP63 gene, belonging to the TP53 superfamily). The Krüppel-like factor 4 (KLF4), repressor element-1 (RE-1)-silencing transcription factor (REST) corepressor 1 (RCOR1), lysine demethylase 1A (KDM1A), and C-terminal-binding protein 1/2 (CTBP1/2) chromatin regulators cooperate with ZNF750 to repress epidermal progenitor genes and activate the expression of epidermal terminal differentiation genes (Sen et al., 2012; Boxer et al., 2014). Besides, ZNF750 and the regulatory network composed of bone morphogenetic protein (BMP) signaling pathway, long non-coding RNAs (lncRNAs) (anti-differentiation non-coding RNA (ANCR) and tissue differentiation-inducing non-protein coding RNA (TINCR)), musculoaponeurotic fibrosarcoma oncogene (MAF)/MAF family B (MAFB), grainy head-like 3 (GRHL3), and positive regulatory domain zinc finger protein 1 (PRDM1) jointly promote epidermal cell differentiation (Sen et al., 2012).
Adenocarcinoma/metabolism*
;
Carcinogenesis/genetics*
;
Colonic Neoplasms/metabolism*
;
Histone Demethylases/metabolism*
;
Humans
;
RNA, Long Noncoding/genetics*
;
Transcription Factors/metabolism*
;
Tumor Suppressor Proteins/metabolism*
5.ICSI causes abnormal H3K9 methylation in the male pronuclei and growth retardation of mouse embryos.
Na TANG ; Xiao-Hong WANG ; Xin-Xin LIANG ; Jian-Bo LI ; Dong WANG ; Yuan LIU ; Bo LI
National Journal of Andrology 2013;19(7):593-598
OBJECTIVETo evaluate the safety of intracytoplasmic sperm injection (ICSI) in the mouse model.
METHODSWe simulated clinical ICSI technology and comprehensively evaluated it by parthenogenetic activation, immunofluorescence, embryo transplantation, examination of early implantation, and measurement of the crown-rump length (CRL).
RESULTSICSI significantly reduced the ability of preimplantation embryo development of the mouse, especially after the 8-cell stage (P < 0.01). The fluorescence of H3K9 dimethylation was abnormal at the male pronuclei of the embryos derived from ICSI. Further examination of the development of the transferred ICSI embryos indicated no significant difference in the rate of early implantation at E5. 5 days as compared with normal fertilization (P = 0.6), but the percentage of "normal embryos" was decreased significantly at E9.5 days (P < 0.01). Obvious growth retardation phenotype was observed even in the normal ICSI embryos at E9.5 days.
CONCLUSIONICSI might result in growth retardation of embryos by affecting H3K9 dimethylation in the male pronuclei.
Animals ; Embryonic Development ; Female ; Histones ; metabolism ; Jumonji Domain-Containing Histone Demethylases ; metabolism ; Male ; Methylation ; Mice ; Mice, Inbred ICR ; Pregnancy ; Sperm Injections, Intracytoplasmic ; adverse effects
6.Effect of Morus alba extract sanggenon C on growth and proliferation of glioblastoma cells.
Wen-Han TANG ; Zhi-Ning ZHANG ; Hua-Rui CAI ; Wei SUN ; He YANG ; Er-Hu ZHAO ; Hong-Juan CUI
China Journal of Chinese Materia Medica 2023;48(1):211-219
Glioblastoma is the most common primary cranial malignancy, and chemotherapy remains an important tool for its treatment. Sanggenon C(San C), a class of natural flavonoids extracted from Morus plants, is a potential antitumor herbal monomer. In this study, the effect of San C on the growth and proliferation of glioblastoma cells was examined by methyl thiazolyl tetrazolium(MTT) assay and 5-bromodeoxyuridinc(BrdU) labeling assay. The effect of San C on the tumor cell cycle was examined by flow cytometry, and the effect of San C on clone formation and self-renewal ability of tumor cells was examined by soft agar assay. Western blot and bioinformatics analysis were used to investigate the mechanism of the antitumor activity of San C. In the presence of San C, the MTT assay showed that San C significantly inhibited the growth and proliferation of tumor cells in a dose and time-dependent manner. BrdU labeling assay showed that San C significantly attenuated the DNA replication activity in the nucleus of tumor cells. Flow cytometry confirmed that San C blocked the cell cycle of tumor cells in G_0/G_1 phase. The soft agar clone formation assay revealed that San C significantly attenuated the clone formation and self-renewal ability of tumor cells. The gene set enrichment analysis(GSEA) implied that San C inhibited the tumor cell division cycle by affecting the myelocytomatosis viral oncogene(MYC) signaling pathway. Western blot assay revealed that San C inhibited the expression of cyclin through the regulation of the MYC signaling pathway by lysine demethylase 4B(KDM4B), which ultimately inhibited the growth and proliferation of glioblastoma cells and self-renewal. In conclusion, San C exhibits the potential antitumor activity by targeting the KDM4B-MYC axis to inhibit glioblastoma cell growth, proliferation, and self-renewal.
Humans
;
Glioblastoma/genetics*
;
Bromodeoxyuridine/therapeutic use*
;
Signal Transduction
;
Proto-Oncogene Proteins c-myc/metabolism*
;
Agar
;
Cell Proliferation
;
Cell Line, Tumor
;
Apoptosis
;
Jumonji Domain-Containing Histone Demethylases/metabolism*
7.Lysine-specific demethylase 1 represses THP-1 monocyte-to-macrophage differentiation.
Rui-feng YANG ; Guo-wei ZHAO ; Shu-ting LIANG ; Hou-zao CHEN ; De-pei LIU
Chinese Medical Sciences Journal 2013;28(2):82-87
OBJECTIVETo investigate the role of lysine-specific demethylase 1 (LSD1) in the process of THP-1 monocyte-to-macrophage differentiation.
METHODSQuantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were performed to analyze the expression of LSD1 and interleukin-6 (IL-6) in THP-1 monocytes and THP-1-derived macrophages. Chromatin immunoprecipitation (ChIP) assay was applied to detect the occupancy of LSD1 and H3K4 methylation at IL-6 promoter during THP-1 monocyte-to-macrophage differentiation. IL-6 mRNA level and H3K4 methylation at IL-6 promoter were analyzed using qRT-PCR and ChIP assay in LSD1-knockdown THP-1 cells treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 0, 4, 8, 12, and 24 hours. Fluorescence activated flow cytometry was performed to reveal the percentage of macrophages differentiated from THP-1 monocytes.
RESULTSThe expression of LSD1 reduced during THP-1 monocyte-to-macrophage differentiation (P<0.01). LSD1 occupancy decreased and H3K4 methylation increased at IL-6 promoter during the differentiation. With knockdown of LSD1, H3K4 methylation at IL-6 promoter was found increased after TPA treatment at different times points (all P<0.05, except 24 hours). The percentage of macrophages increased significantly in the THP-1 cells with LSD1 knockdown (P<0.05).
CONCLUSIONSLSD1 is repressed during the monocyte-to-macrophage differentiation of THP-1 cells. Suppression of LSD1-mediated H3K4 demethylation may be required for THP-1 monocyte-to-macrophage differentiation.
Cell Differentiation ; Cells, Cultured ; Dealkylation ; Histone Demethylases ; physiology ; Histones ; metabolism ; Humans ; Interleukin-6 ; genetics ; Macrophages ; cytology ; Monocytes ; cytology ; Promoter Regions, Genetic
8.Antiproliferative effect of silencing LSD1 gene on Jurkat cell line and its mechanism.
Shiwei HAN ; Yiqun HUANG ; Ruiji ZHENG
Chinese Journal of Hematology 2016;37(1):56-60
OBJECTIVETo investigate the effect of silencing LSD1 gene by RNA interference on the proliferation, apoptosis on human lymphocytic leukemia Jurkat cell line and its mechanism.
METHODSThe hairpin- like oligonucleotide sequences targeting LSD1 gene was transfected into Jurkat cells by lipofectamine(TM) 2000. The LSD1 mRNA and protein were detected by RQ- PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expression of Bcl-2, Bax, procaspase- 3, and histone H3K4me, H3K4me2, H3K4me3, Act- H3, H3K9me were detected by Western blot.
RESULTSLSD1 mRNA was markedly suppressed by the shRNA targeting LSD1. LSD1 shRNA suppressed the proliferation and induced cells apoptosis of Jurkat cells. The cell apoptotic rate was (41.34±3.58)%, (3.45±1.54)%, (1.76±0.52)% in LSD1 shRNA, Neg-shRNA and Blank respectively, the difference among them was statistically significant (P<0.05). LSD1 shRNA down- regulated the expressions of Bcl- 2 and procaspase- 3, and up- regulated the expression of Bax. The methylation of H3K4me1, me2 and acetylation of Act- H3 improved without change of the methylation of H3K4me3.
CONCLUSIONSDeplete of LSD1 gene maybe through modifying the methylation of histone H3K4 to promote the cell apoptosis and inhibit cell growth in Jurkat cell line.
Acetylation ; Apoptosis ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Histone Demethylases ; genetics ; Histones ; metabolism ; Humans ; Jurkat Cells ; Methylation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Transfection
9.Histone demethylase JMJD3 downregulation protects against aberrant force-induced osteoarthritis through epigenetic control of NR4A1.
Yu JIN ; Zhen LIU ; Zhenxia LI ; Hairui LI ; Cheng ZHU ; Ruomei LI ; Ting ZHOU ; Bing FANG
International Journal of Oral Science 2022;14(1):34-34
Osteoarthritis (OA) is a prevalent joint disease with no effective treatment strategies. Aberrant mechanical stimuli was demonstrated to be an essential factor for OA pathogenesis. Although multiple studies have detected potential regulatory mechanisms underlying OA and have concentrated on developing novel treatment strategies, the epigenetic control of OA remains unclear. Histone demethylase JMJD3 has been reported to mediate multiple physiological and pathological processes, including cell differentiation, proliferation, autophagy, and apoptosis. However, the regulation of JMJD3 in aberrant force-related OA and its mediatory effect on disease progression are still unknown. In this work, we confirmed the upregulation of JMJD3 in aberrant force-induced cartilage injury in vitro and in vivo. Functionally, inhibition of JMJD3 by its inhibitor, GSK-J4, or downregulation of JMJD3 by adenovirus infection of sh-JMJD3 could alleviate the aberrant force-induced chondrocyte injury. Mechanistic investigation illustrated that aberrant force induces JMJD3 expression and then demethylates H3K27me3 at the NR4A1 promoter to promote its expression. Further experiments indicated that NR4A1 can regulate chondrocyte apoptosis, cartilage degeneration, extracellular matrix degradation, and inflammatory responses. In vivo, anterior cruciate ligament transection (ACLT) was performed to construct an OA model, and the therapeutic effect of GSK-J4 was validated. More importantly, we adopted a peptide-siRNA nanoplatform to deliver si-JMJD3 into articular cartilage, and the severity of joint degeneration was remarkably mitigated. Taken together, our findings demonstrated that JMJD3 is flow-responsive and epigenetically regulates OA progression. Our work provides evidences for JMJD3 inhibition as an innovative epigenetic therapy approach for joint diseases by utilizing p5RHH-siRNA nanocomplexes.
Cartilage, Articular/pathology*
;
Chondrocytes/metabolism*
;
Down-Regulation
;
Epigenesis, Genetic
;
Humans
;
Jumonji Domain-Containing Histone Demethylases/metabolism*
;
Nuclear Receptor Subfamily 4, Group A, Member 1/metabolism*
;
Osteoarthritis/pathology*
;
RNA, Small Interfering/pharmacology*
10.Effect of Jumonji domain-containing protein-3 on the proliferation and migration of lung cancer cell line.
Cuijie TIAN ; Hongyu DENG ; Xiaoju TANG ; Hefang HU ; Xiaojing LIU ; Fengming LUO
Journal of Biomedical Engineering 2012;29(3):514-518
For investigating the effect of Jumonji domain-containing protein-3 (JMJD3) on the behavior of lung cancer cell line, A549 proliferation was measured with EDU staining and flow cytometer after JMJD3 expression plasmid and pcDNA3. 1 transfection at 48h. The migration ability of A549 was tested at the same time. The expression of p21 mRNA was measured with RT-PCR. The results showed that JMJD3 transfection increased the EDU positive cells ratio (JMJD3: 40.75% +/- 2.07%, control: 20.97% +/- 1.5%, P < 0.001). G1 phase cell ration also increased after JMJD3 transfection (JMJD3:47. 80% +/- 1.85%, control: 54.60% +/- 0.95%, P = 0.005). The mRNA expression of p21 decreased in JMJD3 group (JMJD3: 35. 89% +/- 3.71%, control: 91.78% +/- 3.74%, P < 0.001). The distances of migration were (0.47 +/- 0.27) cm and (0.96 +/- 0.40) cm after 24h and 48h with JMJD3 tranfection, compared to (0.57 +/- 0.22)cm and (1.08 +/- 0.33)cm in control, respectively (P > 0.05). JMJD3 promoted the proliferation of A549 and decreased the G1 cell numbers, decreased the p21 mRNA, but had no effect on A549 migration.
Adenocarcinoma
;
pathology
;
Cell Line, Tumor
;
Cell Movement
;
drug effects
;
Cell Proliferation
;
drug effects
;
Cyclin-Dependent Kinase Inhibitor p21
;
genetics
;
metabolism
;
Humans
;
Jumonji Domain-Containing Histone Demethylases
;
pharmacology
;
Lung Neoplasms
;
pathology
;
RNA, Messenger
;
genetics
;
metabolism
;
Transfection