Effects and related mechanisms of histone deacetylase inhibitors on the proliferation of choroidal melanoma C918 cell line / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To elucidate the effect of histone deacetylase(HDAC)inhibitor suberoylanilide hydroxamic acid(SAHA)on the proliferation of choroidal melanoma(CM)cell line C918 and to explore the related mechanism.METHODS: Inverted fluorescence microscope was used to observe the effect of different concentrations of SAHA(0.625, 1.25 or 2.5 μmol/L)on the morphology of C918 cell. The cell viability was detected by cholecystokinin octapeptide(CCK-8)assay. Plate clone formation assay and EdU staining were carried out to measure the effect of SAHA on the cell proliferation. Meanwhile, the expressions of cell proliferation-related proteins including c-Myc, CyclinA2 and CDK2, and histone deacetylase 7(HDAC7)and fibroblast growth factor 18(FGF18)were detected by Western blot.RESULTS: Compared with the control group, the cell density was reduced in SAHA. SAHA could also promote cell shrinkage, and the inhibition on cell was in a concentration-dependent manner. CCK-8 assay showed that SAHA treatment decreased cell viability in a dose-dependent manner and the inhibition rate was 80% when SAHA at 2.5 μmol/L. Compared with the control group, Western blot showed that SAHA could suppress the expression of cell proliferation proteins including c-Myc, CyclinA2 and CDK2 in a dose-dependent manner. In addition, 1.25 μmol/L SAHA significantly decreased the numbers of EdU staining positive cells and cell clones. More importantly, SAHA could dose-dependently decrease the expression of HDAC7 and FGF18 compared with control group.CONCLUSION: SAHA could inhibit the proliferation of CM cell line C918 by inhibiting the HDAC7/FGF18 signaling pathway.

In vivo inhibitory effect of suberoylanilide hydroxamic acid combined with sorafenib on human hepatocellular carcinoma cells

The present study aimed to investigate the in vivo inhibitory effect of histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) combined with sorafenib on human hepatocellular carcinoma HCCLM3 cells. The nude mice transplanted with HCCLM3 cells were randomly divided into control, SAHA, sorafenib and SAHA+sorafenib groups. The nude mice in the later 3 groups were intragastrically administrated with SAHA (10 mg·kg-1·day-1), sorafenib (10 mg·kg-1·day-1) and SAHA (10 mg·kg-1·day-1) combined with sorafenib (10 mg·kg-1·day-1), respectively, for successive 20 days. Finally, the inhibition rate of tumor was measured. The expressions of MEK1/2, p-ERK1/2, Cyclin D1, Bcl-2, Bax, p53, MMP-2, MMP-9 and uPA in tumor tissues were determined. Results showed that, compared with SAHA and Sorafenib groups, in SAHA+sorafenib groups the inhibition rate of tumor was significantly increased (P < 0.05), the expression levels of MEK1/2, p-ERK1/2, Cyclin D1, Bcl-2, MMP-2 and MMP-9 and uPA protein in tumor tissues were significantly decreased, respectively (P < 0.05), and the expression levels of Bax and p53 protein were significantly increased, respectively (P < 0.05). In conclusion, compared with single drug, SAHA combined with sorafenib can enhance the inhibitory effects on HCCLM3 xenografts in nude mice.

Effect and mechanism of suberoylanilide hydroxamic acid on fear extinction in mice with chronic so-cial defeat stress / 中华行为医学与脑科学杂志

Objective To investigate the effect and mechanism of suberoylanilide hydroxamic acid (SAHA) on the fear extinction in mice with chronic social defeat stress (SD). Methods Fifty-six male C57BL/6J mice aged 7-8 weeks were randomly divided into control group,social defeat group,control-SAHA group and social defeat-SAHA group to investigate the effect of SAHA and social defeat group,social defeat-AAV BDNF group and social defeat-AAV blank group to investigate the effect of BDNF. Fear extinction in mice was evaluated by fear conditioning test (FC). The levels of BDNF and HDAC2 in mice hippocampus were detected by Western blot (WB). The expression of BDNF-overexpressing virus in hippocampus of mice was detected by immunofluorescence assay. Results (1) Compared with control group,fear extinction in the social defeat group was significantly decreased (P<0. 05). Compared with control group, the level of HDAC2(0. 50±0. 02) in the social defeat group was significantly increased (P<0. 001),while the level of BDNF(0. 16 ± 0. 03) was significantly decreased (P<0. 001) in the social defeat group. ( 2) After using SAHA,fear extinction of mice significantly improved (P<0. 05). Compared with control group,the level of HDAC2 (0. 26±0. 02) in the control-SAHA group was significantly decreased(P<0. 001),and the level of BDNF (0. 40±0. 03) was significantly increased (P<0. 001). Compared with social defeat group,the level of HDAC2 (0. 39±0. 03) in the social defeat-SAHA group was significantly decreased (P<0. 001),and the lev-el of BDNF (0. 28±0. 01) was significantly increased (P<0. 001). (3)After injection BDNF-overexpressing virus,fear extinction was significantly improved(P<0. 05). Conclusion SAHA can enhance fear extinction in mice with chronic social defeat stress and its mechanism may be related to the up-regulation of BDNF ex-pression in hippocampus by inhibiting HDAC2 in hippocampal.

Enhancement of histone deacetylase inhibitors SAHA in chemotherapeutic sensitivity of ovarian cancer SKOV3 cells / 吉林大学学报(医学版)

Objective: To investigate the biological effects of histone deacetylase inhibitors (HDACIs) suberoylanilide hydroxamic acid (SAHA) combined with vincristine (VCR) on the ovarian cancer SKOV3 cells, and to clarify their mechanisms. Methods: The human ovarian cancer SKOV3 cells were divided into control group, SAHA group, VCR group and SAHA+VCR group. CCK-8 method was used to detect the surival rates of SKOV3 cells in various groups. The apoptotic rates, the autophagy rates and the distribution of cell cycles of SKV03 cells in various groups were detected by flow cytometry (FCM). Results: The CCK-8 results showed that compared with control group, the survival rates of SKOV3 cells in SAHA group and VCR group were decreased significantly. Compared with VCR group or SAHA group, the survival rates of cells in SAHA + VCR groups were decreased significantly (P

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.

High-affinity Binding Sites for Estrogen Receptor α in the p21WAF1/CIP1 Promoter Region in Breast Cancer MCF-7 Cells / 中国医科大学学报

Objective To investigate the specific sites that estrogen receptor (ER)α could be recruited to in the p21WAF1/CIP1 promoter region to regulate its transcriptional activity in MCF-7 cells,and to clarify the molecular mechanism of suberoylanilide hydroxamic acid (SAHA) and leptin in the regulation of p21WAFI/CIP1 promoter function.Methods MCF-7 cells were starved by culturing them in fetal calf serum-free medium for 24 hours,and then treated with 20 μmol/L of 0.88 μL SAHA (SAHA group) or 0.625 nmol/L of 10 μL leptin (leptin group) for 24 hours,or cultured in complete RPMI-1640 medium (control group).Cell lysates were incubated with anti-ERα antibody for ChIP analysis.The relative expression levels of DNA fragments,ranging from the TSS to upstream of the p21WAF1/CIP1 promoter region (+2 to-4 000 bp),that bound the antibody were detected by real-time PCR.Results In the control group,the relative expression levels of f1,f2,and f8 DNA fragments that bound the antiERα antibody were two-fold higher than the relative expression of the f9 fragment (P ＜ 0.01).In the SAHA and leptin groups,the relative expression of f1 to f10 DNA fragment that bound anti-ERα antibody was significantly lower than that of the control.The binding affinity of ERα for the f8 fragment was the lowest (P ＜ 0.01) in the SAHA group,and it was significantly lower than that in the leptin group (P ＜ 0.01).Conclusion ERα could be recruited to the p21WAFI/CIP1 promoter via signaling pathways activated during the proliferation of breast cancer MCF-7 cells.Moreover,the DNA fragment ranging from-2 800 to-3 200 bp upstream of the p21 WAF1/CIP1 promoter is the target functional region for high-affinity binding with ERα.

Synergistic effect of ERα on HDAC1 regulatingtranscription of p21WAF1/CIP1 in breast cancer MCF-7 cells / 中国药理学通报

Aim To investigate the specific binding sites that histone deacetylases 1(HDAC1) and estrogen receptor α(ERα)can be recruited to regulate the transcriptional activity of p21WAF1/CIP1 promoter in the breast cancer MCF-7 cells, and to clarify the molecular mechanism of suberoylanilide hydroxamic acid(SAHA) and leptin regulating p21WAF1/CIP1 promoter function.Methods The breast cancer MCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours, and treated with 20 μmol·L-1 SAHA(SAHA group) or 0.625 nmol·L-1 leptin(Leptin group) for 24 hours.The cells that were cultured in complete RPMI 1640 medium without any treatment were assigned as control group(Basal group).The cell lysis was prepared and incubated respectively with anti-HDAC1 and anti-ERα antibody by chromatin-immunoprecipitation(ChIP) method overnight at 4℃.The DNA-ChIP was followed the manufacturer′s protocol for the assay.DNA fragments binding anti-HDAC1 and anti-ERα antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21WAF1/CIP1 promoter region(+2~-4 000 bp) binding with antibody was detected by real-time PCR and analyzed by 2-△△CT method.Results In basal group, HDAC1 and ERα had high affinity with the f1 and f8 fragments of p21WAF1/CIP1 promoter compared to the f4 fragment.In SAHA group, the binding ability of HDAC1 and ERα to the f1 and f8 fragments of p21WAF1/CIP1 promoter was significantly lower than that of the control, while reversing to reach the peak after leptin treatment.Conclusions HDAC1 and ERα can be recruited to p21WAF1/CIP1 promoter by the cell proliferation signal during the proliferation of breast cancer MCF-7 cells.The DNA f1(from 0 to-400 bp) and f8(from-2 800 to-3 200 bp) fragment in the upstream of p21WAF1/CIP1 promoter are the target functional region for the binding with HDAC1and ERα.

Suberoylanilide hydroxamic acid induces apoptosis of rat hepatic stellate cells in vitro / 中国病理生理杂志

AIM:To study the effects of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hepatic stellate cells (HSCs) and expression of associated proteins, and to investigate the mechanisms of SAHA to induce apoptosis.METHODS:The rat HSCs were isolated by OptiPrep gradient centrifugation method.The effect of SAHA on HSC proliferation was detected by real-time cell analyzer.The morphological changes of HSCs treated with SAHA at different concentrations were observed under inverted microscope.The apoptotic rates of HSCs were analyzed by flow cytometry with Annexin V-FITC/PI staining and fluorescence microscopy.The protein expression of α-smooth muscle actin (α-SMA), collagen I, tissue inhibitor of metalloproteinase 1 (TIMP1), glucose-regulated protein 78 (GRP78) and histone deacetylase 6 (HDAC6) was detected by Western blotting.The interaction of GRP78 with HDAC6 in the HSCs was determined by co-immunoprecipitation.RESULTS:HSCs were successfully isolated and cultured for 14 d, during which the HSCs changed gradually from rest state to active state.SAHA significantly inhibited the proliferation of HSCs in a time-and dose-dependent manner (P<0.05).The results of Western blotting showed that the protein expression levels of α-SMA, TIMP1, collagen-I and HDAC6 were significantly decreased (P<0.05), while GRP78 was significantly increased (P<0.05).Compared with activated HSCs, GRP78 and total acetyl-lysine protein were significantly increased in the co-immunoprecipitated HSCs treated with SAHA, while HDAC6 protein was significantly decreased, indicting that GRP78 formed a complex with HDAC6.CONCLUSION:The anti-hepatic fibrosis effect of SAHA may be related to down-regulation of HDAC6 and up-regulation of acetylated GRP78, thus inducing endoplasmic reticulum stress of HSCs and promoting the apoptosis of HSCs.

Effects of suberoylanilide hydroxamic acidon proliferation and apoptosis of human hepatic stellate cells / 基础医学与临床

Objective To determine the effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid(SAHA) on the cell proliferation and apoptosis of the human hepatic stellate cell line LX-2.The possible underlying mechanisms were also investigated.Methods The LX-2 cells were treated with SAHA in vitro.The morphology of LX-2 cells in different concentrations groups was observed by inverted microscope;the proliferation of LX-2 cells was measured by MTT assay;the Annexin V-FITC and PI staining was used to detect the apoptosis of LX-2 cells by flow cytometry and fluorescence microscope;the expression of α-SMA,collagen Ⅰ,acH3K9,acH3K14 and acH3K18 were detected by Western blot.Results The morphology change of LX-2 cells showed that SAHA inhibited the proliferation rate of LX-2 cells and in a dose dependent manner(P<0.05).The LX-2 cells were sensitive to SAHA along with time increasing,and in a time-dependent manner(P<0.05).Western blot showed that the expression levels of α-SMA and collagen-Ⅰ were significantly lower(P<0.05),on the contrary,the acetylation levels of acH3K9,acH3K14 and acH3K18 were significantly higher (P<0.05).Conclusions The increased acetylation of the histone acH3K9,acH3K14,acH3K18 and the lower expressed α-SMA and collagen-Ⅰ in LX-2 cells may be one of the mechanisms of SAHA.

A study on high function binding site of HDAC1 in p21 WAF1/CIP1 promoter region in breast cancer MCF-7 cells / 中国药理学通报

Aim Toinvestigatethespecificbinding sites that HDAC1 can be recruited to regulate the tran-scriptional activity of p21 WAF1/CIP1 promoter in the breast cancer MCF-7 cells.Methods ThebreastcancerMCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours,and treated with 20 μmol·L-1 SAHA(S group)or 0. 625 nmol·L-1 Leptin(L group)for 24 hours,and the cells that were cultured in the complete RPMI 1640 medium without any treatment were assigned as control group (B group).The DNA-ChIP was followed the manufactur-er′s protocol for the assay.The cell lysis was prepared and incubated with anti-HDAC1 antibody overnight at 4℃.DNA fragments binding anti-HDAC1 antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21 WAF1/CIP1 promoter region(+2 ~-4000 bp)bind-ing with antibody was detected by Real-time PCR and analyzedby2-ΔΔCTmethod.Results InBgroup, HDAC1 had high affinity with the f1 and f 8 fragmentsof p21 WAF1/CIP1 promoter compared to the other fragemts,and showed the highest affinity with the f8 fragment.In S group,the binding ability of HDAC1 to the f1 ~f10 fragment of p21 WAF1/CIP1 promoter was sig-nificantly lower than that of the control.The binding activity of HDAC1 to f8 fragment was the lowest,while reversing to reach the peak after leptin treatment.Con-clusions HDAC1canberecruitedtop21WAF1/CIP1pro-moter by the cell proliferation signal during the prolifer-ation of breast cancer MCF-7 cells.The DNA fragment from -2800 to -3200 bp in the upstream of p21 WAF1/CIP1 promoter is the target functional region for the binding with HDAC1 .

Suberoylanilide hydroxamic acid induces apoptosis of HepG2 cells by en-doplasmic reticulum stress apoptotic pathway / 中国病理生理杂志

AIM:To investigate the effect of suberoylanilide hydroxamic acid ( SAHA) on the proliferation and apoptosis of human hepatocellular carcinoma HepG 2 cells and to explore its possible mechanism .METHODS: HepG2 cells were treated with SAHA at different concentrations for 48 h.The proliferation of HepG2 cells was detected by real-time cellular analysis.The protein levels of acetylated histones H3K9 and H3K27, glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase ( PERK ) and p-PERK were determined by Western blot .The cell apoptosis was analyzed by flow cytometry .RESULTS:Compared with control group , treatment with SAHA at 0.1μmol/L and 1 μmol/L for 48 h showed no significant inhibitory effect on the proliferation of HepG 2 cells, while SAHA at 6 μmol/L and 12 μmol/L significantly inhibited the proliferation of HepG 2 cells (P<0.05).The results of Western blot showed that the protein levels of acH3K9, acH3K27, GRP78 and p-PERK increased significantly after treated with SAHA at diffe-rent concentrations for 48 h, while the protein level of PERK was decreased significantly (P<0.05).The results of flow cytometry analysis showed that the apoptotic rates of the HepG 2 cells increased with the increase in SAHA concentration . CONCLUSION:SAHA up-regulates the acetylation of H3K9 and H3K27 in the HepG2 cells and induces apoptosis of HepG2 cells by activating the endoplasmic reticulum stress-related apoptosis pathway .

Effects and the possible mechanism of histone deacetylase inhibitor SAHA on the interstitial fibrosis induced by diabetes / 中华肾脏病杂志

Objective To explore the effects and possible mechanism of histone deacetylase inhibitor SAHA on the interstitial fibrosis induced by diabetes.Methods The SD rats were divided into three groups:control group (Con,n=9),diabetes mellitus (DM) group (n=9) and SAHA treatment group (n=9).The diabetic rat model was established by injecting streptozotocin (STZ) through tail vein.After 8 weeks,the SAHA treatment group rats were fed with a SAHA solution (25 mg· kg-1 · d-1) by gastric gavage.After 16 weeks,all rats were sacrificed to detect relevant biochemical parameters,and observe the changes of pathomorphology in kidney.In addition,immunohistochemistry staining and Western blotting were employed to detect the protein expressions of transforming growth factor-β1 (TGF-β1),Smad2,Smad3,p-Smad2,p-Smad3,Smad7,collagen-Ⅰ and collagen-Ⅲ,respectively.Results Compared with Con group,the levels of blood glucose (BG),urinary trace albumin/urinary creatinine (ACR),triglyceride (TG) and total cholesterol (TC) in the diabetic group were all increased significantly (all P ＜ 0.05),the protein expressions of TGF-β1,p-Smad2,p-Smad3,collagen-Ⅰ and collagen-Ⅲ in kidney were all increased in diabetic group (all P ＜ 0.05),and the expression of Smad7 was significantly reduced (P ＜ 0.05).Compared with DM group,the levels of ACR was reduced,the renal fibrosis was alleviated,the protein expressions of TGF-β1,p-Smad2,p-Smad3,collagen-Ⅰ and collagen-Ⅲ in SAHA group were all decreased (all P ＜ 0.05),and the expression of Smad7 was increased significantly (P ＜ 0.05).Conclusion SAHA may restore the protein level of Smad7 by enhancing protein stability,then promote the moderate transduction of TGF-β1/Smads signaling pathway,which reduce the fibrosis of renal tubules in diabetic rats.

Notch3 pathway mediates SAHA-induced apoptosis in human small-cell lung cancer H446 cells / 中国病理生理杂志

AIM: To investigate the effect of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hu-man small-cell lung cancer H446 cells and its possible mechanism.METHODS: H446 cells were incubated in the medi-um containing SAHA.CCK-8 assay was used to detect the anti-tumor effect of SAHA on the H446 cells, and IC50 values of SAHA were calculated.Flow cytometry was used to analyze the apoptosis.After Notch3 gene was silenced, the pro-apopto-tic effect of SAHA on the H446 cells was inhibited ( P <0.05).Eukaryotic expression plasmid containing N3ICD was transfected into the H446 cells, so that N3ICD was expressed in the H446 cells.The mRNA expression of Notch3 was measured by RT-PCR.The protein levels of Notch3, N3ICD, Puma and cleaved caspase-3 were determined by Western blot.RESULTS: SAHA remarkably reduced the cell viability in a dose-dependent manner (P <0.05), and the IC50 value of SAHA was 1.91 μmol/L.SAHA induced apoptosis in a dose-dependent manner (P <0.05).The expression of Notch3 gene was negative in the H446 cells, SAHA reactivated Notch3 gene and Notch3 pathway in a dose-dependent manner (P <0.05).Notch3 knockdown inhibited apoptosis induced by SAHA (P <0.05).Over-expression of N3ICD up-regula-ted the protein levels of Puma and cleaved caspase-3.CONCLUSION: SAHA induces apoptosis in human small-cell lung cancer H446 cells by activating Notch3 pathway and up-regulating the protein level of Puma.

Effects of Combination of SAHA and TRAIL on Proliferation of Triple-negative Breast Cancer Cell MDA-MB-231 / 中国医科大学学报

Objective To investigate the effects of combined treatment of suberoylanilide hydroxamic acid(SAHA)and TNF?related apoptosis inducing ligand(TRAIL)on proliferation and morphology change of triple?negative breast cancer cell MDA?MB?231. Methods The effects of combination treatment of SAHA and TRAIL on proliferation and morphology change of MDA?MB?231 cells were monitored by RTCA. Morphology changes of MDA?MB?231 cells by different treatment factors were observed through time?lapse live cell imaging acquisition. Results Real?time cell proliferation assays showed that a synergistic effect were found when MDA?MB?231 cells were treated with combination of SAHA and TRAIL , and reached the best effect with 5μmol/L SAHA and 50 ng/mL TRAIL. The results of time?lapse live cell imaging acquisition showed that the growth inhibition of MDA?MB?231 cells with combined treatment of SAHA and TRAIL were more obvious than that with treatment of SAHA or TRAIL alone. Conclusion The combined treatment of SAHA and TRAIL induces a synergistic effect on growth inhibition in triple?negative breast cancer cell line MDA?MB?231.

The inhibitory effects of suberoylanilide hydroxamic acid on the malignant phenotypes of ovarian carcinoma cells / 天津医药

Objective To explore the effects and molecular mechanisms of suberoylanilide hydroxamic acid (SAHA) on ovarian carcinoma. Methods (1)Two groups of ovarian carcinoma cell lines (SKOV3 and SKOV3/DDP, HO8910 and HO8910-PM) were exposed to SAHA (1, 3, 5 and 7μmol/L SAHA,group 1-group 4). CCK-8 method was employed to eval-uate the inhibitory effects of SAHA.(2)Ovarian cancer cell lines treated with SAHA (2 or 5μmol/L SAHA) were used as 1 and 2 groups. Flow cytometry was performed following staining with Annexin V-FITC and PI for cell cycle and apoptosis.(3) Reverse transcription polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the mRNA and pro-tein expression levels of phenotypic correlation factor. Results (1)After 48 h of SAHA treatment,the OD value of SKOV3, SKOV3/DDP,and HO8910 showed a trend of gradually reduce (P<0.05).(2)The apoptotic rates were significantly higher in SAHA 1 and SAHA 2 groups than those of control group (P<0.05). Compared with control group, after 48 h of SAHA treat-ment,S phase and G2/M phase of SKOV3 and SKOV3/DDP cells increased;G0/G1 phase of HO8910 and HO8910-PM cells increased in SAHA 1 and 2 groups (P<0.05).(3)The expression levels of CyclinB1 and Cdc2 (p34) mRNA were significant-ly lower in SAHA 1 and 2 groups than those of control group,while the expression levels of Caspase-3,p21 and p53 mRNA expression were significantly higher in SAHA 1 and 2 groups than those of control group. Furthermore,the expression of Ac-Histone H3,Ac-Histone H4,p53 protein were markedly improved,and CyclinB1,Cdc2(p34) protein decreased in SAHA 1-4 groups. Conclusion SAHA may suppress cell growth, induce apoptosis and cause cycle arrest in ovarian carcinoma cells by promoting histone acetylation or modulating their phenotype-related proteins of Caspase-3, p53, CyclinB1 and Cdc2(p34).

Effects of combination with SAHA and TRAIL treatment on cell growth of er positive breast cancer cell line MCF-7 / 中国药学杂志

OBJECTIVE: To study the effects of suberoylanilide hydroxamic acid (SAHA) and TRAIL treatment on cell proliferation and apoptosis for ER positive breast cancer cell line MCF-7. METHODS: Human breast cell lines (MCF-7) were evaluated for the expressions of cell viability, cell apoptosis and cell cycle by muse cell analyzer. The mRNA levels of related apoptotic factors in MCF-7 cells were detected by real time PCR and solid phase apoptosis antibody microarray. RESULTS: After the combination with SAHA and TRAIL, the ability of cell proliferation and cell viability were depressed, and the cell apoptosis was induced. The cell cycle assay showed that the MCF-7 cells were arrested in G0/G1 phase with SAHA and TRAIL treatment. CONCLUSION: The combinatorial treatment of SAHA and TRAIL has a significantly inhibitory effect on cell growths of ER positive breast cancer MCF-7 cell.

SAHA inhibition of the human ovarian cancer cell line SKOV3 in vitro / 中国肿瘤临床

10.3969/j.issn.1000-8179.2013.12.004

Suberoylanilide hydroxamic acid (SAHA) suppresses JAK/STAT1 signal pathway to down-regulate the expression of indoleamine 2, 3-dioxygenuse in gallbladder carcinoma cells / 中华肝胆外科杂志

Objective To investigate the mechanism on JAK/STAT1 signal pathway in SAHA down-regulation of indoleamine 2,3-dioxygenase (IDO) in gallbladder carcinoma cells.MethodsWe treated gallbladder carcinoma SGC-996 cells with IFN-γ and SAHA.Western blotting was used to detect the expression of IDO,signal transducer and activator of transcription 1 (STAT1) phosphorylation and interferon regulatory factor genes-1 (IRF-1).Confocal microscopy analysis was used to detect STAT1 translocation.Transient transfections and reporter genes assay was used in detecting the activation of γ-activated sites (GAS) and interferon stimulated response elements (ISRE).ResultsIDO expressed in SGC-996 cells in dose- and time-dependent manners when stimulated with IFN-γ.SAHA down-regulated the expression of IDO induced by IFN-γ in a dose-dependent manner.SAHA blocked the expression of IRF-1 induced by IFN-γ.SAHA inhibited the IFN-γ-induced STAT1 phosphorylation and nuclear translocation.SAHA down-regulated IFN-γ-induced activation of GAS and ISRE.ConclusionsSAHA may down-regulate IDO expression through inhibiting the activation of members in JAK/STAT1 signal pathway.This may provide a new immunotherapeutic strategy to break tumor immune tolerance in gallbladder carcinoma.

The effect of combined treatment with cisplatin and histone deacetylase inhibitors on HeLa cells / 부인종양

OBJECTIVE: To investigate the combined effects of cisplatin and the histone deacetylase (HDAC) inhibitors suberoylanilide hydroxamic acid (SAHA) or sirtinol on HeLa cells and assess the mechanism underlying HDAC inhibitor-cisplatin synergy. METHODS: The antineoplastic actions of cisplatin, SAHA and sirtinol, alone and in combination, were evaluated using the tetrazolium dye-based MTT cell proliferation assay, DAPI nuclear staining and cytotoxicity analysis. RESULTS: Exposure to cisplatin, SAHA or sirtinol alone induced a dose-dependent reduction in HeLa cell viability. Combined treatment with cisplatin and SAHA or sirtinol was significantly more cytotoxic than cisplatin alone. Individually, cisplatin, SAHA and sirtinol activated caspase-3 and induced apoptosis, but the effects of combined treatment were greater. Importantly, both HDAC inhibitors dose-dependently inhibited the expression of the antiapoptotic proteins Bcl-2 and x-linked inhibitor of apoptosis protein (XIAP). CONCLUSION: The combination of cisplatin and SAHA or sirtinol had synergistic effect on the HeLa cell viability. This potentiation of cisplatin activity was associated with HDAC inhibitor-mediated down-regulation of Bcl-2 and XIAP. These may result from the relaxation of chromatin by these HDAC inhibitors that increase cisplatin sensitivity by enhancing the accessibility of DNA to cisplatin and transcriptional regulators.