1.Role and mechanism of histone deacetylases in mouse neuronal development.
Yu-Wei BAI ; Meng-Long GUAN ; Tao ZHENG ; Shi-Ping LI ; Yi QU ; De-Zhi MU
Chinese Journal of Contemporary Pediatrics 2021;23(3):294-299
OBJECTIVE:
To study the role and mechanism of histone deacetylase 1 (HDAC1) and histone deacetylase 2 (HDAC2) in mouse neuronal development.
METHODS:
The mice with Synapsin1-Cre recombinase were bred with
RESULTS:
The mice with
CONCLUSIONS
Deletion of
Animals
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Blotting, Western
;
Histone Deacetylase 1/genetics*
;
Histone Deacetylase 2
;
Histone Deacetylases/genetics*
;
Immunohistochemistry
;
Mice
;
Neurons/metabolism*
;
Signal Transduction
2.Effect of Histone Deacetylase Inhibition on the Expression of Multidrug Resistance-associated Protein 2 in a Human Placental Trophoblast Cell Line.
Hong-Yu DUAN ; Dan MA ; Kai-Yu ZHOU ; ; Tao WANG ; Yi ZHANG ; ; Yi-Fei LI ; Jin-Lin WU ; Yi-Min HUA ; ; Chuan WANG ;
Chinese Medical Journal 2017;130(11):1352-1360
BACKGROUNDPlacental multidrug resistance-associated protein 2 (MRP2), encoded by ABCC2 gene in human, plays a significant role in regulating drugs' transplacental transfer rates. Studies on placental MRP2 regulation could provide more therapeutic targets for individualized and safe pharmacotherapy during pregnancy. Currently, the roles of epigenetic mechanisms in regulating placental drug transporters are still unclear. This study aimed to investigate the effect of histone deacetylases (HDACs) inhibition on MRP2 expression in the placental trophoblast cell line and to explore whether HDAC1/2/3 are preliminarily involved in this process.
METHODSThe human choriocarcinoma-derived trophoblast cell line (Bewo cells) was treated with the HDAC inhibitors-trichostatin A (TSA) at different concentration gradients of 0.5, 1.0, 3.0, and 5.0 μmol/L. Cells were harvested after 24 and 48 h treatment. Small interfering RNA (siRNA) specific for HDAC1/HDAC2/HDAC3 or control siRNA was transfected into cells. Total HDAC activity was detected by colorimetric assay kits. HDAC1/2/3/ABCC2 messenger RNA (mRNA) and protein expressions were determined by real-time quantitative polymerase chain reaction and Western-blot analysis, respectively. Immunofluorescence for MRP2 protein expression was visualized and assessed using an immunofluorescence microscopy and ImageJ software, respectively.
RESULTSTSA could inhibit total HDAC activity and HDAC1/2/3 expression in company with increase of MRP2 expression in Bewo cells. Reduction of HDAC1 protein level was noted after 24 h of TSA incubation at 1.0, 3.0, and 5.0 μmol/L (vs. vehicle group, all P < 0.001), accompanied with dose-dependent induction of MRP2 expression (P = 0.045 for 1.0 μmol/L, P = 0.001 for 3.0 μmol/L, and P < 0.001 for 5.0 μmol/L), whereas no significant differences in MRP2 expression were noted after HDAC2/3 silencing. Fluorescent micrograph images of MRP2 protein were expressed on the cell membrane. The fluorescent intensities of MRP2 in the control, HDAC2, and HDAC3 siRNA-transfected cells were week, and no significant differences were noticed among these three groups (all P > 0.05). However, MRP2 expression was remarkably elevated in HDAC1 siRNA-transfected cells, which displayed an almost 3.19-fold changes in comparison with the control siRNA-transfected cells (P < 0.001).
CONCLUSIONSHDACs inhibition could up-regulate placental MRP2 expression in vitro, and HDAC1 was probably to be involved in this process.
Cell Line ; Histone Deacetylase 1 ; metabolism ; Histone Deacetylase 2 ; metabolism ; Histone Deacetylase Inhibitors ; pharmacology ; Histone Deacetylases ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; Microscopy, Fluorescence ; Multidrug Resistance-Associated Proteins ; genetics ; metabolism ; RNA, Messenger ; Trophoblasts ; cytology ; metabolism
3.Sulforaphane, an NRF2 agonist, alleviates ferroptosis in acute liver failure by regulating HDAC6 activity.
Yan-Qiong ZHANG ; Chun-Xia SHI ; Dan-Mei ZHANG ; Lu-Yi ZHANG ; Lu-Wen WANG ; Zuo-Jiong GONG
Journal of Integrative Medicine 2023;21(5):464-473
OBJECTIVE:
Acute liver failure (ALF) is characterized by severe liver dysfunction, rapid progression and high mortality and is difficult to treat. Studies have found that sulforaphane (SFN), a nuclear factor E2-related factor 2 (NRF2) agonist, has anti-inflammatory, antioxidant and anticancer effects, and has certain protective effects on neurodegenerative diseases, cancer and liver fibrosis. This paper aimed to explore the protective effect of SFN in ALF and it possible mechanisms of action.
METHODS:
Lipopolysaccharide and D-galactosamine were used to induce liver injury in vitro and in vivo. NRF2 agonist SFN and histone deacetylase 6 (HDAC6) inhibitor ACY1215 were used to observe the protective effect and possible mechanisms of SFN in ALF, respectively. Cell viability, lactate dehydrogenase (LDH), Fe2+, glutathione (GSH) and malondialdehyde (MDA) were detected. The expression of HDAC6, NRF2, glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and solute carrier family 7 member 11 (SLC7A11) were detected by Western blotting and immunofluorescence.
RESULTS:
Our results show that NRF2 was activated by SFN. LDH, Fe2+, MDA and ACSL4 were downregulated, while GSH, GPX4 and SLC7A11 were upregulated by SFN in vitro and in vivo, indicating the inhibitory effect of SFN on ferroptosis. Additionally, HDAC6 expression was decreased in the SFN group, indicating that SFN could downregulate the expression of HDAC6 in ALF. After using the HDAC6 inhibitor, ACY1215, SFN further reduced HDAC6 expression and inhibited ferroptosis, indicating that SFN may inhibit ferroptosis by regulating HDAC6 activity.
CONCLUSION
SFN has a protective effect on ALF, and the mechanism may include reduction of ferroptosis through the regulation of HDAC6. Please cite this article as: Zhang YQ, Shi CX, Zhang DM, Zhang LY, Wang LW, Gong ZJ. Sulforaphane, an NRF2 agonist, alleviates ferroptosis in acute liver failure by regulating HDAC6 activity. J Integr Med. 2023; 21(5): 464-473.
Humans
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Ferroptosis
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NF-E2-Related Factor 2/genetics*
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Liver Failure, Acute/drug therapy*
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Isothiocyanates/pharmacology*
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Glutathione
;
Histone Deacetylase 6
4.Effect of histone acetylation/deacetylation imbalances on key gene of planar cell polarity pathway.
Hong-Yu DUAN ; Yi ZHANG ; Kai-Yu ZHOU ; Chuan WANG ; DA-Jian QIU ; Yi-Min HUA
Chinese Journal of Contemporary Pediatrics 2017;19(4):475-483
OBJECTIVETo investigate the effect of histone acetylation/deacetylation imbalances on embryonic hearts of mice and its effect on key genes of planar cell polarity (PCP) pathway-Vangl2, Scrib and Rac1 in H9C2 cells.
METHODSForty pregnant C57/B6 mice were randomly assigned into three groups: blank group (n=10), vehicle group (n=10), and valproic acid (VPA)-treated group (n=20). In the VPA-treated group, VPA, a histone deacetylase (HDAC) inhibitor, was administered to each individual dam intraperitoneally at a single dose of 700 mg/kg on embryonic day 10.5 (E10.5). The vehicle and blank groups received equivalent saline or no interventions, respectively. Dams were sacrificed on E15.5, and death rates of embryos were evaluated. Subsequently, embryonic hearts of survival fetus were removed to observe cardiac abnormalities by hematoxylin-eosin (HE) staining. H9C2 cells were cultured and allotted to the blank, vehicle, and VPA-treated groups: the VPA treated group received VPA exposure at concentrations of 2.0, 4.0 and 8.0 mmol/L; the vehicle and blank groups received equivalent saline or no interventions, respectively. HDAC1-3 as well as Vangl2, Scrib and Rac1 mRNA and protein expression levels were determined by quantitative real-time PCR and Western blot, respectively. The total HDAC activity was analyzed by colorimetric assay.
RESULTSThe fetus mortality rate after VPA treatment was 31.7%, with a significantly higher rate of cardiac abnormalities in comparison with the controls (P<0.05). In comparison with the blank and vehicle groups, HDAC1 mRNA was significantly increased at various concentrations of VPA treatment at all time points of exposure (P<0.05), together with a reduction of protein level after 48 and 72 hours of exposure (P<0.05). The inhibition of HDAC2 mRNA after various concentrations of VPA incubation was pronounced at 24 hours of exposure (P<0.05), while the protein levels were reduced at all time points (P<0.05). HDAC3 mRNA was prominently induced by VPA (4.0 and 8.0 mmol/L) at all time points of treatment (P<0.05). In contrast, the protein level was inhibited after VPA treatment (P<0.05). In comparison with the blank and vehicle groups, Vangl2 mRNA as well as Scrib mRNA/protein expression levels were markedly reduced after 48 and 72 hours of VPA treatment (P<0.05), together with a reduction of protein level in Vangl2 at 72 hours (P<0.05). Compared with the blank and vehicle groups, a significant repression in the total HDAC activity was observed in the VPA-treated group at concentrations of 4.0 and 8.0 mmol/L after 24 hours of treatment (P<0.05), and the effect persisted up to 48 and 72 hours, exhibiting pronounced inhibition at all concentrations (P<0.05).
CONCLUSIONSVPA might result in acetylation/deacetylation imbalances by inhibiting HDAC1-3 protein expression and total HDAC activity, leading to the down-regulation of mRNA and protein expression of Vangl2 and Scrib. This could be one of the mechanisms contributing to congenital heart disease.
Acetylation ; Animals ; Cell Polarity ; Cells, Cultured ; Fetal Heart ; drug effects ; metabolism ; Heart Defects, Congenital ; etiology ; Histone Deacetylase 1 ; genetics ; Histone Deacetylase 2 ; genetics ; Histones ; metabolism ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; genetics ; RNA, Messenger ; analysis ; Valproic Acid ; pharmacology
5.Selective class I histone deacetylase inhibitors suppress persistent spontaneous nociception and thermal hypersensitivity in a rat model of bee venom-induced inflammatory pain.
Fan YANG ; Yan YANG ; Yan WANG ; Fei YANG ; Chun-Li LI ; Xiao-Liang WANG ; Zhen LI ; Jun CHEN
Acta Physiologica Sinica 2015;67(5):447-454
To confirm whether class I histone deacetylase inhibitors (HDACIs) are effective in relief of peripheral inflammatory pain, the effects of two selective inhibitors, MS-275 and MGCD0103, were studied in rats inflamed by subcutaneous (s.c.) injection of bee venom (BV). The BV test is characterized by displaying both persistent spontaneous nociception (PSN) and primary hypersensitivity. Intrathecal (i.t.) pre-treatment of either MS-275 or MGCD0103 with a single dose of 60 nmol/20 μL resulted in profound suppression of both PSN and primary thermal hypersensitivity but without significant influence upon the primary mechanical hypersensitivity and mirror-image thermal hypersensitivity. Moreover, the up-regulation of both HDAC1 and HDAC2 induced by s.c. BV injection was completely suppressed by i.t. pre-treatment of MS-275. The present results provide with another new line of evidence showing involvement of epigenetic regulation of chromatin structure by HDAC1/2-mediated histone hypoacetylation in the BV-induced PSN and thermal hypersensitivity and demonstrate the beneficial effects of class I HDACIs in prevention of peripheral inflammatory pain from occurring.
Animals
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Bee Venoms
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administration & dosage
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Benzamides
;
pharmacology
;
Epigenesis, Genetic
;
Histone Deacetylase 1
;
genetics
;
metabolism
;
Histone Deacetylase 2
;
genetics
;
metabolism
;
Histone Deacetylase Inhibitors
;
pharmacology
;
Hot Temperature
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Hyperalgesia
;
drug therapy
;
Inflammation
;
drug therapy
;
Injections, Subcutaneous
;
Nociception
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Pain
;
chemically induced
;
drug therapy
;
Pain Measurement
;
Pyridines
;
pharmacology
;
Pyrimidines
;
pharmacology
;
Rats
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Rats, Sprague-Dawley
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Up-Regulation
6.A polymorphism in the histone deacetylase 1 gene is associated with the response to corticosteroids in asthmatics.
Min Hye KIM ; Sae Hoon KIM ; Yook Keun KIM ; Soo Jong HONG ; Kyung Up MIN ; Sang Heon CHO ; Heung Woo PARK
The Korean Journal of Internal Medicine 2013;28(6):708-714
BACKGROUND/AIMS: Recent investigations suggest that histone deacetylase 1 (HDAC1) and HDAC2 may be target molecules to predict therapeutic responses to corticosteroids. We evaluated the effects of variation in HDAC1 and HDAC2 on the response to corticosteroids in asthmatics. METHODS: Two single nucleotide polymorphisms (SNPs) were selected after resequencing HDAC1 and HDAC2. For the first analysis, we evaluated the association between those SNPs and asthma severity in 477 asthmatics. For the second analysis, we evaluated the effects of these SNPs on lung function improvements in response to corticosteroid treatment in 35 independent adult asthmatics and 70 childhood asthmatics. RESULTS: We found that one SNP in HDAC1 (rs1741981) was significantly related to asthma severity in a recessive model (corrected p = 0.036). Adult asthmatics who were homozygous for the minor allele of rs1741981 showed significantly lower % forced expiratory volume in 1 second (%FEV1) increases in response to systemic corticosteroids treatment compared with the heterozygotes or those homozygous for the major allele (12.7% +/- 7.2% vs. 37.4% +/- 33.7%, p = 0.018). Similarly, childhood asthmatics who were homozygous for the minor allele of rs1741981 showed significantly lower %FEV1 increases in response to inhaled corticosteroid treatment compared with the heterozygotes or those homozygous for the major allele (14.1% +/- 5.9% vs. 19.4% +/- 8.9%, p = 0.035). CONCLUSIONS: The present study demonstrated that rs1741981 in HDAC1 was significantly associated with the response to corticosteroid treatment in asthmatics.
Administration, Inhalation
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Adrenal Cortex Hormones/administration & dosage/*therapeutic use
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Adult
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Aged
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Anti-Asthmatic Agents/administration & dosage/*therapeutic use
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Asthma/diagnosis/*drug therapy/enzymology/genetics/physiopathology
;
Child
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Female
;
Forced Expiratory Volume
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Gene Frequency
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Heterozygote
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Histone Deacetylase 1/*genetics
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Histone Deacetylase 2/genetics
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Homozygote
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Humans
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Lung/*drug effects/physiopathology
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Male
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Middle Aged
;
Pharmacogenetics
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Phenotype
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*Polymorphism, Single Nucleotide
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Recovery of Function
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Severity of Illness Index
;
Treatment Outcome
7.Effect of down-regulation of histone deacetylase 2 protein expression on cell proliferation and cell cycle in cervical carcinoma.
Fang-fang HUA ; Yong-hua XIA ; Da-peng WU ; Rui-xiang CHEN ; Yu-hong WANG ; Ying PAN ; Jun YANG ; Wu-feng LIANG
Chinese Journal of Pathology 2012;41(7):466-469
OBJECTIVETo study the effect of down-regulation of histone deacetylase 2 (HDAC2) expression on cell proliferation and cell cycle in cervical carcinoma cell lines HeLa.
METHODSHDAC2 siRNA and control siRNA were transfected to HeLa cells. CCK-8 and flow cytometry were used to analyze the changes of cell proliferation and cell cycle, respectively. Western blot was employed to detect the changes of cell proliferation and cell cycle-related proteins.
RESULTSHDAC2 siRNA significantly down-regulated the expression of HDAC2 protein in HeLa cells, resulting in marked inhibition of cell proliferation. In addition, the percentage of cells in G(0)/G(1) phase in HDAC2 siRNA group (63.3% ± 2.0%) was significantly higher than that in untreated group (29.3% ± 1.7%) or control siRNA group (29.4% ± 1.7%), F = 354.181, P = 0.000. Furthermore, Western blot demonstrated that down-regulation of HDAC2 expression decreased the expression of cyclin D1, cyclin E and CDK2 proteins but increased the expression of p21 protein.
CONCLUSIONSDown-regulation of HDAC2 expression mediates proliferation inhibition and cell cycle arrest. It is associated with decrease in cyclin D1, cyclin E and CDK2 protein expression and increase in p21 protein expression.
Cell Cycle ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase 2 ; metabolism ; Down-Regulation ; HeLa Cells ; Histone Deacetylase 2 ; genetics ; metabolism ; Humans ; Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; RNA, Small Interfering ; genetics ; Transfection
8.Effect of HDAC6 down-regulation on the growth of xenografted human laryngeal carcinoma cell line Hep-2 in nude mice and underlying mechanism.
Lin-lin YUAN ; Wei-hua LOU ; Jian-zhong SANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(6):481-486
OBJECTIVETo study the effect of histone deacetylation 6 (HDAC6) siRNA on the growth of xenografted human laryngeal squamous cell carcinoma cell line Hep-2 in nude mice and underlying mechanism.
METHODSLaryngeal squamous cell carcinoma cell line Hep-2 cells were subcutaneously injected to the back of nude mice and transplanted tumor model was established after one week. Nude mice was divided into three groups including blank control group, empty vector group and HDAC6 siRNA group, and the tumor growth was observed. Ki-67 proliferation index was detected by immunohistochemistry. Western blot, in situ hybridization and immunohistochemistry were used to detect the mRNA and protein expressions of HDAC6 in xenograft. The expressions of Bcl-2 and Bax proteins were examined by Western blotting. Cell apoptosis was detected by TUNEL.
RESULTSThe mean volume of xenograft transfected with HDAC6 siRNA was less than that of xenograft transfected with empty vector or that of xenograft with blank control treatment (P < 0.05). HDAC6 siRNA effectively down-regulated the expressions of HDAC6 mRNA and the expressions of HDAC6 and Bcl-2 proteins, but up-regulated the expression of Bcl-2 protein in xenografts, with significant differences (all P < 0.05). The proliferation index of Ki-67 in HDAC6 siRNA transfection group was significantly lower than that in blank control group or empty vector group (P < 0.05). TUNEL assay demonstrated that HDAC6 evidently evoked cell apoptosis (P < 0.05).
CONCLUSIONHDAC6 siRNA could effectively inhibited the growth of xenografted human laryngeal carcinoma cell line Hep-2 in nude mice, down-regulate the expressions of HDAC6 and bcl-2, and up-regulate the expression of bax.
Animals ; Cell Line, Tumor ; Down-Regulation ; Female ; Histone Deacetylase 6 ; Histone Deacetylases ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism
9.ATM modulates transcription in response to histone deacetylase inhibition as part of its DNA damage response.
Eun Ryoung JANG ; Jae Duk CHOI ; Mi Ae PARK ; Gajin JEONG ; Hyeseong CHO ; Jong Soo LEE
Experimental & Molecular Medicine 2010;42(3):195-204
Chromatin structure has a crucial role in a diversity of physiological processes, including development, differentiation and stress responses, via regulation of transcription, DNA replication and DNA damage repair. Histone deacetylase (HDAC) inhibitors regulate chromatin structure and activate the DNA damage checkpoint pathway involving Ataxia-telangiectasia mutated (ATM). Herein, we investigated the impact of histone acetylation/deacetylation modification on the ATM-mediated transcriptional modulation to provide a better understanding of the transcriptional function of ATM. The prototype HDAC inhibitor trichostain A (TSA) reprograms expression of the myeloid cell leukemia-1 (MCL1) and Gadd45alpha genes via the ATM-mediated signal pathway. Transcription of MCL1 and Gadd45alpha is enhanced following TSA treatment in ATM+ cells, but not in isogenic ATM- or kinase-dead ATM expressing cells, in the ATM-activated E2F1 or BRCA1-dependent manner, respectively. These findings suggest that ATM and its kinase activity are essential for the TSA-induced regulation of gene expression. In summary, ATM controls the transcriptional upregulation of MCL1 and Gadd45alpha through the activation of the ATM-mediated signal pathway in response to HDAC inhibition. These findings are important in helping to design combinatory treatment schedules for anticancer radio- or chemo-therapy with HDAC inhibitors.
Cell Cycle Proteins/genetics/*metabolism
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DNA Damage/*genetics
;
DNA-Binding Proteins/*metabolism
;
E2F1 Transcription Factor/metabolism
;
Gene Expression Regulation/drug effects
;
Histone Deacetylase Inhibitors/*pharmacology
;
Histone Deacetylases/*metabolism
;
Humans
;
Hydroxamic Acids/pharmacology
;
Nuclear Proteins/genetics/metabolism
;
Promoter Regions, Genetic/genetics
;
Protein Binding/drug effects
;
Protein-Serine-Threonine Kinases/*metabolism
;
Proto-Oncogene Proteins c-bcl-2/genetics/metabolism
;
RNA, Messenger/genetics/metabolism
;
*Transcription, Genetic/drug effects
;
Tumor Suppressor Proteins/*metabolism
10.Down-regulation of histone deacetylase 2 induces cell apoptosis and inhibits cell proliferation and migration of laryngeal squamous cell carcinoma cells.
Lin-lin YUAN ; Wei-hua LOU ; Jian-zhong SANG ; Sheng-lei LI ; Wen-cai LI
Chinese Journal of Pathology 2012;41(6):396-399
OBJECTIVETo investigate the effects of histone deacetylase 2 (HDAC2) expression on cell proliferation, apoptosis and migration of laryngeal squamous cell carcinoma (LSCC) Hep-2 cells.
METHODSHDAC2 siRNA and control siRNA were transfected into LSCC Hep-2 cells by lipofectamine 2000, and cells were divided into three experimental groups: untreated group, control siRNA group and HDAC2 siRNA transfection group. Western blotting was utilized to detect the expression of HDAC2 protein in Hep-2 cells. Cell proliferation and apoptosis were investigated by CCK-8 kit and flow cytometry, respectively. Boyden chamber was used to study cell migration. Expressions of cell apoptosis and cell migration related proteins were detected by Western blotting.
RESULTSHDAC2 siRNA significantly down-regulated the expression of HDAC2 protein in LSCC Hep-2 cells. Down-regulation of HDAC2 expression coincided with an inhibition of cell proliferation and migration along with an induced cell apoptosis of Hep-2 cells. Moreover, down-regulation of HDAC2 expression significantly increased the expressions of caspase-3 and caspase-9 proteins but decreased the expressions of matrix metalloproteinases (MMP)-2 and MMP-9 proteins.
CONCLUSIONSHDAC2 may play a pivotal role in the initiation and development of LSCC. Down-regulation of HDAC2 expression mediates cell apoptosis. Cell migration inhibition may be tightly associated with overexpression of caspase-3 and caspase-9 along with down-regulation of MMP-2 and MMP-9 expressions.
Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Histone Deacetylase 2 ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection