Effects and mechanism of ginsenoside Rh2 on the proliferation and apoptosis of human glioma cells / 中国药房

ABSTRACT OBJECTIVE To investigate the effects and mechanism of ginsenoside Rh2 on the proliferation and apoptosis in human glioma U87 and U251 cells. METHODS Using human glioma U87 and U251 cells as subjects， the proliferation and apoptosis， as well as the expression of histone deacetylase 1（HDAC1） protein and apoptosis-related proteins [B cell lymphoma-2（Bcl-2）， Bcl-2-associated X protein （Bax） and cleaved caspase-3] were detected after being treated with different concentrations of ginsenoside Rh2. RESULTS The concentrations of 10，20，30，40，50，60，70，80 μmol/L ginsenoside Rh2 could generally significantly increase the proliferation inhibition rate of U87 and U251 cells （P＜0.05 or P＜0.01）， and the half inhibitory concentrations of this component after 48 hours of action were 51.34 and 55.84 μmol/L， respectively；30，50 μmol/L ginsenoside Rh2 could increase the total apoptotic rate of both types of cells， reduced the protein expressions of HDAC1 and Bcl-2， and increased the protein expressions of Bax and cleaved caspase-3 significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Ginsenoside Rh2 has a significant inhibitory effect on the proliferation of glioma cells and promotes the apoptosis of cells， which may be through reducing the expression of HDAC1 protein and activating the Bcl-2 family protein-mediated apoptosis pathway.

Influence of histone deacetylase 1 on insulin resistance in a cell model of nonalcoholic fatty liver disease / 临床肝胆病杂志

Objective To investigate the promoting effect of histone deacetylase 1 (HDAC1) expression on insulin resistance (IR) in nonalcoholic fatty liver disease (NAFLD) cells by establishing an HepG2 cell model of high fat-induced NAFLD. Methods HepG2 cells were divided into control group, model group (OA), and inhibitor group (OA+pyroxamide [an HDAC1 inhibitor]). CCK-8 assay was used to plot the standard growth curve of HepG2 cells and screen out the optimal drug concentration and action time of OA and pyroxamide; oil red O staining was used to compare the accumulation of lipid droplets in cells; an automatic biochemical analyzer was used to analyze the content of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), and total cholesterol (TC) in cells; quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of HDAC1 and insulin receptor substrate-1 (IRS-1) in cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results OA treatment at a concentration of 0.25 mmol/L for 24 hours was the optimal concentration and duration of cell modeling, and treatment at a concentration of 20 μmol/L for 24 hours was the optimal administration concentration and duration of pyroxamide. Compared with the control group, the model group had significant increases in the content of ALT, AST, TG, and TC, and compared with the model group, the inhibitor group had significant reductions in the content of ALT, AST, TG, and TC (all P < 0.05). The model group had significantly higher mRNA and protein expression levels of HDAC1 than the control group, while the inhibitor group had significantly lower expression levels than the model group (all P < 0.05); the model group had significantly lower mRNA and protein expression levels of IRS-1 than the control group, while the inhibitor group had significantly higher expression levels than the model group (all P < 0.05). Conclusion HDAC1 participates in the development and progression of NAFLD by inhibiting the expression of IRS-1 molecule and promoting IR, and the HDAC1 inhibitor pyroxamide can exert a protective effect on the liver by alleviating IR.

Early intervention of bone-nearby acupuncture combined with electroacupuncture on morphine tolerance in bone cancer pain rats and its effect on the expression of HDAC and MOR in dorsal root ganglia / 中国针灸

OBJECTIVE@#To observe the effect of early intervention of bone-nearby acupuncture (BNA) combined with electroacupuncture (EA) on the expression of histone deacetylase1(HDAC1), histone deacetylase 2 (HDAC2) andμ-opioid recepter (MOR) in dorsal root ganglia (DRG) of bone cancer pain-morphine tolerance (BCP-MT) rats, and to explore its possible mechanism.@*METHODS@#A total of 35 SD rats were randomized into a sham BCP group (=6), a BCP group (=7), a MT group (=7), a BNA+EA group (=8) and a shame BNA group (=7). Except of the sham BCP group, cancer cell inoculation operation at left tibia was given in the other 4 groups to establish the bone cancer pain model. In the MT group, the BNA+EA group and the shame BNA group, intraperitoneal injection of morphine hydrochloride was given to establish the morphine tolerance model. After the operation, bone-nearby acupuncture combined with electroacupuncture was applied at "Zusanli" (ST 36) and "Kunlun" (BL 60) in the BNA+EA group, with dilatational wave, 2 Hz/100 Hz in frequency, 0.5 to 1.5 mA in intensity. Intervention in the shame BNA group was applied at the same time and acupoints as those in the BNA+EA group, the needles were pierced the skin without any electrical stimulation. The needles were retained for 30 min, once a day for continuous 7 days in both BNA+EA and shame BNA groups. Before and 10, 11, 15, 22 days after the operation, the left paw withdrawal threshold (PWT) was measured in the 5 groups. The levels of HDAC1, HDAC2 and MOR in DRG were detected by Western blot.@*RESULTS@#Ten days after the cancer cell inoculation operation, the PWT of the BCP, MT, BNA+EA and sham BNA groups was decreased compared with the sham BCP group (0.05); the PWT of the BNA+EA group was increased compared with the MT and sham BNA group (<0.01). In the BCP group, the DRG levels of HDAC1 and HDCA2 were increased, while the level of MOR was decreased compared with the sham BCP group (<0.05, <0.01). In the MT group, the DRG level of HDAC1 was increased compared with the BCP group (<0.05). In the BNA+EA group, the DRG level of HDAC1 was decreased compared with the MT group and the sham BNA group (<0.01, <0.05), while the level of MOR was increased (<0.01).@*CONCLUSION@#Early intervention of bone-nearby acupuncture combined with electroacupuncture can relieve the morphine tolerance in bone cancer pain rats, it may relate to down-regulating the expression of HDAC1 and up-regulating the expression of MOR in the dorsal root ganglia.

Effect of HDAC1 silencing on apoptosis of squamous cell carcinoma of skin via regulating STAT3 signaling pathway / 中国病理生理杂志

AIM:To investigate the effect of histone deacetylase 1(HDAC1)silencing on apoptosis of squa-mous cell carcinoma of skin.METHODS:Skin squamous cell carcinoma A431 cells were transfected with HDAC1 small interfering RNA(HDAC1 siRNA)or small interfering RNA negative control(siRNA NC).The expression levels of HDAC1 in transfected cells were detected by RT-PCR and Western blot.The cell viability was measured by MTT assay, and the apoptosis was analyzed by flow cytometry.The protein levels of STAT3,p-STAT3 and cleaved caspase-3 were de-termined by Western blot.The inhibitor of STAT3 signaling pathway was used to treat the A 431 cells transfected with HDAC1 siRNA.The cell viability was detected by MTT assay,the apoptosis was analyzed by flow cytometry,and the pro-tein levels of STAT3,p-STAT3 and cleaved caspase-3 were determined by Western blot.RESULTS: HDAC1 siRNA in-hibited the expression of HDAC1 at mRNA and protein levels in the A431 cells.After interfering with the expression of HDAC1,the cell viability and the protein level of p-STAT3 in the cells decreased,while the apoptotic rate and the protein level of cleaved caspase-3 in the cells were increased.After treatment with the inhibitor of STAT3 pathway,the viability of A431 cells transfected with siRNA and the protein level of p-STAT3 decreased,while the apoptotic rate and the protein le-vel of cleaved caspase-3 in the cells were increased.CONCLUSION: Interference with HDAC1 expression may regulate the STAT3 signaling pathway to inhibit the viability of skin squamous cell carcinoma cells,thus promoting the apoptosis of squamous cell carcinoma of skin.

Effects of SAHA on Twist and HDAC1 expression in renal tissues of dia-betic rats / 中国病理生理杂志

AIM:To observe the effects of suberoylanilide hydroxamic acid(SAHA )on the expression of Twist,histone deacetylase 1(HDAC1),and the factors associated with epithelial-mesenchymal transition and extracellular matrix in diabetic rat renal tissues and its possible mechanism.METHODS:The rat model of diabetes mellitus(DM)was established by injection of streptozotocin.The rats were randomly divided into normal control(NC)group,DM group and SAHA group.When the model was successfully established for 8 weeks ,the rats in SAHA group were intragastrically trea-ted with SAHA at dose of 25 mg· kg-1 · d-1.After 8 weeks of treatment ,the rats were sacrificed.The biochemical pa-rameters and renal index were measured ,and the pathomorphological changes of the renal tissues were observed by HE and Masson staining.In addition ,the expression of Twist in renal tubular epithelial cells was evaluated by immunohistochemis -try.The protein levels of Twist,HDAC1,E-cadherin,α-SMA and collagen type Ⅳ(Col-Ⅳ)were determined by Western blot.The mRNA expression of Twist was detected by real-time PCR.RESULTS:The blood glucose ,24 h urine protein and kidney index in DM group were all obviously higher than those in NC group ,while kidney index was reduced in SAHA group as compared with DM group(P<0.05),but the blood glucose and 24 h urine protein markedly wasn't influenced by SAHA treatment.Compared with NC group,the mRNA and protein levels of Twist and protein levels of HDAC 1,α-SMA and Col-Ⅳwere increased in DM group(P<0.05),while they decreased in SAHA group as compared with DM group(P<0.05).The protein level of E-cadherin in NC group was higher than that in DM group ,and that was increased in SAHA group as compared with DM group(P<0.05).The correlation analysis showed that the Twist protein level was positively correlated with the HDAC1 protein level(P<0.05).CONCLUSION:SAHA decreases the expression of Twist and attenuates renal tubule fibrosis in DM rats by inhibiting the expression of HDAC 1 and Twist,thus promoting the tran-scription of E-cadherin.

Changes and clinical significance of peripheral blood HDAC1 and VEGF expression in patients with lung adenocarcinoma / 重庆医学

Objective To investigate the correlation between histone deacetylase 1(HDAC1) and vascular endothelial growth factor(VEGF) in the patients with lung adenocarcinoma.Methods Eighty cases of lung adenocarcinoma in our hospital from August 2014 to April 2016 served as the research subjects,and contemporaneous 80 individuals undergoing healthy physical examination were taken as the control group.The fasting venous blood sample was collected in all subjects.Then serum HDAC1 and VEGF levels were detected by ELISA.The differences of serum HDAC1 and VEGF expression levels were compared between the two groups.The HDAC1 and VEGF expression levels in the patients with different characteristics of lung adenocarcinoma and the relation between serum HDAC1 and VEGF concentrations were analyzed.Furthermore the possible influence factors of HDAC1 protein expression level in the patients with lung adenocarcinoma were analyzed.Results The HDAC1 levels in the control group and observation group were(329.56 ± 23.83) ng/L and(568.20 ± 35.40) ng/L,the difference was statistically significant(t=23.576,P=0.000).The VEGF levels in the control group and observation group were(40.26±9.82)ng/L and(296.56±19.80)ng/L respec tively,the difference was statistically significant(t=31.154,P=0.000).The HDAC1 protein level had statistical difference among different genders,ages,clinical stages and smoking history,the HDAC1 protein level in male,age ＞60 years old,clinical stage Ⅲ,V and patients with smoking history were higher(P＜0.05).The Pearson correlation analysis results showed that serum HDAC1 in the patients with lung adenocarcinoma was positively correlated with VEGF protein concentration(r=0.526,P =0.000).The Logistic regression analysis showed that influence factor of HDAC1 protein expression level in the patients with lung adenocarcinoma was clinical stage.Conclusion The high expression of HDAC1 protein in lung adenocarcinoma patients may also simultaneously regulate the VEGF expression,thus promotes the development of lung adenocarcinoma.

HMGB1 increases radiosensitivity by interacting with HDAC1 / 中华放射医学与防护杂志

Objective To study the nuclear protein association of high-mobility group box-1 (HMGB1) and histone deacetylase 1 (HDAC1),and the effect of interaction on radiosensitivity in human breast cancer cells.Methods The protein-protein interaction was determined by immunoprecipitationWestern blot and glutathione-S-transferase capture assays.Cell growth was examined by MTT (methyl thiazolyl tetrazolium)assay and clonogenic assay.Histone deacetylase activity was analyzed by histone deacetylase assay.Results A significant increase of HMGB1 protein and radiosensitivity was observed in MDA-MB-231 and MDA-MB-468 cells transfected with a pCMV-Tag2B expression vector carrying with a full-length of HMGB1 cDNA.HMGB1 binding to HDAC1 was demonstrated as GST (glutathione Stransferase)-pull down and immunoprecipitation Western blot assay,and the association was elevated by irradiation.An LXCXE motif was required for the HMGB1-HADC1 interaction and HMGB1 radiosensitization.A significant difference of IC50 value was observed,for example,1.8 and 2.2 Gy (wtHMGB1 transfectants,P ＜ 0.05),3.6 and 3.8 Gy (HMGB1/C103F transfectants,P ＞ 0.05),both compared with 3.9 and 4.1 Gy (pCMV-Tag2B transfectants) in MDA-MB-231 and MDA-MB-468 cells,respectively.A specific HDAC1 inhibitor trichostatin A markedly reduced the HMGB1-mediated radiosensitivity,0.5 Gy in the presence of trichostatin A versus 1.8 Gy in absence of trichostatin A in MDA-MB-231 transfectants,1.2 Gy (with trichostatin A) versus 2.2 Gy (without trichostatin A) in MDA-MB-468 transfectants,P ＜ 0.05.Histone deacetylase activity was also detected in immunoprecipitates prepared from these cells with antibodies to HMGB1,and this activity was abolished by the histone trichostatin A.Conclusions These results suggest a previous unanticipated role for HDAC1 in modification of HMGB1-mediated radiosensitivity by its direct interaction with HMGB1.

Association between Expression of HDAC1 Protein and HPV16/18 Infection in Cervical Intraepithelial Neoplasia and Cervical Squamous Cell Carcinoma / 中国医科大学学报

Objective To explore the anomalous expression of HDAC1 in cervical intraepithelial neoplasia(CIN)and cervical carcinoma and its relationship with the infection of HPV16/18. Methods The expressions of HDAC1 protein and HDAC1 infection were detected by in situ hybridiza?tion(ISH)and immunohistochemistry staining(SP)in tissues from patient with chronic cervicitis(the chronic cervicitis group),CIN(the CINⅠgroup,the CINⅡgroup and the CINⅢgroup)and cervical squamous carcinoma(the cervical carcinoma group). Results The infection rate of HPV16/18 was 8.33%,34.78%,48.00%,65.21%,78.00%in patients with chronic cervicitis,CINⅠ,CINⅡ,CINⅢand cervical carcinoma re?spectively. There was significant difference between groups(P<0.05). The positive expression rate of HDAC1 was 0,13.04%,32.00%,47.82%and 76.00%in chronic cervicitis,CINⅠ,CINⅡ,CINⅢand cervical carcinoma,respectively. The differences were statistically significant(P<0.05). The positive expression of HDAC1 in the HPV16/18 infected patients was significantly higher than those uninfected with HPV16/18(P<0.05). The HDAC1 positive expression had no relationship with clinical stages of cervical carcinoma,however,it was related to the tissue differentia?tion,stromal invasion and pelvic lymph node metastasis(all P<0.05). Conclusion The infection of HPV16/18 may play an important role in the carcinongenesis of SCC through increasing the expression of HDAC1.

A polymorphism in the histone deacetylase 1 gene is associated with the response to corticosteroids in asthmatics

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.

HDAC1 Expression in Invasive Ductal Carcinoma of the Breast and Its Value as a Good Prognostic Factor

BACKGROUND: Histone deacetylase 1 (HDAC1) is associated with the expression and function of estrogen receptors and the proliferation of tumor cells, and has been considered a very important factor in breast tumor progression and prognosis. Several studies have reported an association between HDAC1 expression and poorer prognosis in cancers including breast cancer, with a few exceptions. However, because of the dearth of studies on HDAC1 expression in breast cancer, its significance for breast cancer prognosis has not been well defined. Therefore, we examined HDAC1 expression in invasive ductal carcinoma (IDC), the most common breast cancer, and investigated its potential prognostic significance. METHODS: We used 203 IDC tissue samples. Immunohistochemical stains for HDAC1 and real-time polymerase chain reaction for HDAC1 mRNA were performed and the results were compared to generally well-established prognostic factors in breast cancer and patient survival rates. RESULTS: HDAC1 expression was significantly reduced in proportion to higher histologic grade, higher nuclear pleomorphism score, and higher mitotic counts, and with lower estrogen receptor expression. Furthermore, it was significantly associated with the survival rate. CONCLUSIONS: HDAC1 expression is a good prognostic indicator in IDC.

HDAC1 Expression and Effect of Curcumin on Proliferation of Raji Cells / 华中科技大学学报（医学）（英德文版）

Histone deacetylase (HDAC1) has a high expression in many cancer cells and curcumin can inhibit the growth of cancer cells. This paper was designed to investigate the expression of HDAC1 of Raji cells and the effect of curcumin on their proliferation and apoptosis. Raji cells were treated with 3. 125-50 μmol/L curcumin for 8-48 h and the growth inhibition rates of Raji cells were measured by MTT. The expression of HDAC1 on Raji cells were examined by mRNA, Western blot at 24 h various concertrations (1.6-50 μmol/L). Curcumin could selectively inhibit the proliferation of Raji cells in a dose- and time-dependent manner, with the inhibition rate being 52.47 %-82.18%(P＜0.01).The up-regulation of HDAC1 expression was observed within 24 h after the treatment with curcumin as shown by RT-PCR and Western blot. With the increase of concentration, the expression was down-regulated in a dose- dependent manner. It is concluded that the expression of HDAC1 plays an important role in the proliferation and apoptosis of Raji cells and curcumin can inhibit the growth of Raji cells at various concentrations and promote the apoptosis of Raji cells.