SAHA-CTSV induced excessive autophagy and inhibited growth of breast cancer MCF-7 cells / 中国药理学通报

Aim To investigate the mechanism of SA-HA-CTSV inducing autophagy and inhibiting the growth of breast cancer MCF-7 cells. Methods Muse cell analyzer was used to screen the optional treatment doses of drug for cells, and real-time PCR and Western blot were to detect the mRNA and protein expressions of CTSV. Then, specific siRNA was transfected in cells for silencing the CTSV function. Immunofluorescent assay was used to detect the distribution of LC3II, and related ATG molecule expressions were assessed. Bafilomycin A1 was used to inhibit autophagy flux, then p62 protein expression and cell viability were detected. Results 5 μmol · L

Inhibitory effects and mechanism of SAHA combined with Erlotinib on EGFR-TKI resistant lung cancer cell line H1975 cells / 安徽医科大学学报

Objective To investigate the inhibition and possible mechanism of SAHA alone and combined with Er-lotinib on EGFR-TKI resistant lung cancer cell line H1975 cells. Methods The inhibitory effects of SAHA, Erlo-tinib and combined effect on cells were evaluated by CCK-8, colony formation assay and Transwell invasion assay. Western blot analysis was used to analyze the expression of PI3K, AKT, p-AKT, mTOR and p-mTOR protein in cells. Results SAHA and Erlotinib showed growth inhibition effects on H1975 cells with a dose-dependent man-nerand displayed synergistic inhibition effect. The combination of the drugs was significantly stronger than the single drug in the formation and invasion inhibition of tumor cells. The combined group had a strong inhibitory effect on the PI3K/AKT/mTOR signaling pathway. Conclusion SAHA combined with Erlotinib has synergistic inhibitory effects on the growth, cloning and invasion of EGFR-TKI lung cancer cell line H1975, which may be associated with inhibitory effects on the PI3K/AKT/mTOR signaling pathway.

SAHA affects cell cycle of MCF-7 breast cancer cells byregulating acetylated levels of p21WAF1/CIP1 promoter / 中国药理学通报

Aim To study the regulation mechanisms of deacetylase inhibitor SAHA in p21WAF1/CIP1 promoter acetylation in breast cancer MCF-7 cells.Methods We used quantitative real-time PCR, Western blot and DNA-ChIP to determine the effects on the regulation of cell cycle with SAHA treatment in MCF-7 cells.By DNA-ChIP, we assessed the acetylation level of p21WAF1/CIP1 promoter.Results SAHA significantly affected the expression of cell cycle-related factors, and induced the mRNA and protein expression of p21WAF1/CIP1.SAHA could adjust the acetylation level of p21WAF1/CIP1 promoter.Conclusion SAHA regulates the cell cycle progression by adjusting the acetylation level of p21WAF1/CIP1 promoter in MCF-7 cells.

Effects of combination treatment with SAHA and TRAIL on ER positive breast cancer cell MCF-7 / 中国药理学通报

Aim To investigate the effects of com-bined treatment of SAHA and TRAIL on human breast cancer ER positive cell line MCF-7 . Methods MCF-7 cells were treated with SAHA and/or TRAIL. The inhibitory rates were detected by real-time cell prolifer-ation assays. Morphology changes of MCF-7 cells were observed through time-lapse live cell imaging acquisi-tion. Results Real-time cell proliferation assays showed that the anti-tumor efficacy of SAHA was sig-nificantly enhanced in combination with TRAIL. The results of time-lapse live cell imaging acquisition dem-onstrated that, with treatment of SAHA and TRAIL, the growth inhibition of MCF-7 cells was more obvious than that of in TRAIL or SAHA treatment alone. Con-clusion The combination treatment of SAHA and TRAIL has a synergistic effect of growth inhibition on breast cancer MCF-7 cells.

The regulation of Cathepsin B on cell apoptosis induced by SAHA in breast cancer cell line MCF-7 / 中国药理学通报

Aim To clarify the regulation role of ca-thepsin B ( Cat B ) in cell proliferation and apoptosis induced by SAHA in ER-positive breast cancer cell line MCF-7.Methods MTT was used to screen the optimal concentration and treatment time of SAHA . The expression levels of related proteins were deter-mined by ELISA , and the morphological changes were observed through time-lapse live cell imaging acquisi-tion.Cell viability and apoptosis assay in MCF-7 cells were assessed by Muse Cell Analyzer with SAHA and /or Cystatin C treatment .Results MTT assay showed that the anti-tumor efficacy of SAHA was significant . The optimal concentration and treatment time were 10μmol? L-1 and 24 h respectively . ELISA assay showed that SAHA could induce expression of Cat B in MCF-7 cells.Real-time live-cell imaging experiments demonstrated that the combination treatment of Cystatin C and SAHA significantly resumed the inhibitory effect caused by SAHA alone .Cytology test showed that SA-HA alone obviously depressed the cell viability and in-duced apoptosis . However , the effect was reversed with the combination of Cystatin C .Conclusion Cat B plays an important role in apoptosis induced by SAHA in ER +breast cancer cells MCF-7.

SAHA, TRAIL and Apoptosis in Breast Cancer / 沈阳医学院学报

Breast cancer is becoming as the most common malignant tumor that affects women?s health in the world. Numerous stud?ies have shown that histone deacetylase inhibitor SAHA and tumor necrosis factor?related apoptosis?inducing ligand ( TRAIL) can in?duce apoptosis in breast cancer cells, and the combination of TRAIL and SAHA has a synergistic effect, it can significantly enhance the breast cancer cells sensitivity to TRAIL treatment, so the strategy of SAHA and TRAIL treatment on breast cancer has become a welcome and promising method of clinical therapy.

Selection of Chitinolytic Bacteria as Biological Control of Colletotrichum capsici

Aims: The objectives of this study were to screen chitinolytic bacteria isolated from soil of Taman Nasional Bukit Duabelas, Jambi, Indonesia. Isolates were selected based on chitinolytic index and antagonism activity of Colletotrichum capsici. Chitinase enzyme from selected isolates was investigated for growth inhibition of C. capsici. Methodology and results: Two chitinolytic bacteria were selected based on their ability to degrade colloidal chitin and inhibit of the growth of C. capsici. Those isolates were KAHN 15.12 and SAHA 12.12, identified as Serratia marcescens and Bacillus thuringiensis respectively based on 16S rRNA gene. The chitinase maximum specific activity of isolate KAHN 15.12 was 52.03 U/mg after 36 h of incubation and SAHA 12.12 was 45.67 U/mg after 24 h of incubation. The enzyme was precipitated by ammonium sulfate 40% and 60% respectively for KAHN 15.12 and SAHA 12.12. The precipitated chitinases were active over a broad range of pH (5 to 10) and temperature (20 to 80 °C). Enzymes were stable in optimum temperature for 180 min. The precipitated of chitinase KAHN 15.12 and SAHA 12.12 had five and two protein bands respectively on SDS-PAGE gel. Chitinases exhibited an antifungal activity against C. capsici at concentration of 60 ppm. Conclusion, significance and impact of study: Isolates KAHN 15.12 and SAHA 12.12 were successfully selected by their ability to degrade colloidal chitin and inhibit the growth of C. capsici. The isolates had a broad range of pH and temperature, moreover relatively stable at the optimum temperature. Chitinase was effective as biological control for anthracnose caused by C. capsici in chilli.

Regulation of SAHA on cell proliferation induced by leptin in breast cancer cell line MCF-7 / 中国药理学通报

To clarify the molecular mechanism of SAHA in the cell proliferation of ER-positive breast cancer cell line MCF-7 induced by leptin. Methods Human breast cancer cell MCF-7 was incubated with SAHA and/or leptin, and cell viability, apoptosis and cell cy-cle of MCF-7 cells were detected by Muse Cell Analy-zer. The expression of proteins related with apoptosis was determined by apotosis antibody array. Results Real-time cell proliferation assays indicated that the in-duction effect of leptin for MCF-7 cells reached the peak at a concentration of 0. 625 nmol · L-1 . SAHA reduced the viability of MCF-7 cells, induced G0/G1 phase arrest in the cell cycle, and triggered the apopto-sis. Meanwhile, SAHA significantly induced the pro-tein expressions of some apoptotic factors, including Bax, Caspase-3, TRAIL DR5, p21CIP1, and inhibited the expressions of Claspin, Clusterin, x-linked inhibi-tor of apoptosis protein(XIAP) and survivin. Howev-er, leptin had reverse effects on the related expression of the proteins. Conclusion The effects of cell prolif-eration by leptin and SAHA treatment in breast cancer ER positive cell line MCF-7 may involve in the activa-tion of apoptosis pathway, in particular with releasing of Caspase-3 trigged by endogenous mitochondrial ap-optosis pathway.

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.

Effects of SAHA combined with PTX on lethality and autophagy of human ovarian cancer OC3 cells / 军事医学

Objective To evaluate the effect of suberoylanilide hydroxamic acid(SAHA) or/and paclitaxel(PTX) on lethality and autophagy of human ovarian cancer OC3 cells,and to explore whether the combination of the two drugs has a synergistic function.Methods The morphology of OC3 cells was treated with SAHA and/or PTX, and then the morphology of treated OC3 cells was observed under an inverted microscope, cell proliferation was detected by MTT assay and autoph-agy was analyzed by AO/EB double staining assay.The synergistic effect of SAHA and/or PTX was analyzed by factorial design and gold formula method.Results After treatment with SAHA and/or PTX, the morphology of OC3 cells in the combination group ( SAHA+PTX) displayed significant morphological changes.OC3 cells became less adherent and refrac-tive than in other groups.Cell proliferation by MTT assay demonstrated that the growth inhibition rate of the combination groups was higher than in groups treated with SAHA or PTX respectively( P<0.05) .Furthermore, the synergistic effect af-ter treatment with a combination of SAHA with PTX was proved by the factorial design and gold formula method.The auto-phagy rate of the combined groups was significantly higher than in single treatment groups (P<0.05) by AO/EB double staining.Conclusion SAHA and PTX can inhibit the survival of OC3 cells and induce its autophagy.The two drugs have synergistic antitumor effects.

Uso de inibidores de Histona Deacetilase como estratégia terapêutica para sensibilizar células-tronco tumorais a quimioterapia: uma nova visão terapêutica sobre carcinomas mucoepidermoide bucais / Sensitizing Mucoepidermoid Carcinomas to chemotherapy by disrupting the population of Cancer Stem Cells using HDAC inhibitors

Carcinoma mucoepidermóide (CME) é o tumor maligno de glândulas salivares mais comum, representando cerca de 30% dos tumores malignos. O tratamento do CME é a ressecção cirúrgica com eventual radioterapia. Assim, o tratamento do CME pode levar a varias complicações estéticas e funcionais. A quimioterapia tem sido utilizadas apenas em casos recorrentes ou com metástases à distancia. Vários relatos na literatura tem mostrado que o tratamento com drogas isoladas ou combinadas possuem uma resposta insatisfatória e de curta duração em grande parte devido a aquisição de resistência a quimioterapia. Recentemente, a quimiorresistência tem sido relacionada com a presença de Células-Tronco Tumorais (CTT). Essa resistência tem sido associada ao fato de que as essa células são quiescentes e possuem altos níveis de proteínas associadas ao reparo do DNA e baixos níveis das proteínas que levam a apoptose. Recentemente mostrou-se que a resistência a quimioterapia tem sido relacionada com modificações de histonas, uma vez que as células quimiorresistentes possuem núcleo pequeno e baixos níveis de acetilação de histonas, adicionalmente as células sensíveis são relacionadas com núcleo aumentado. O objetivo desse estudo foi avaliar os efeitos do tratamento com IHDAC e cisplatina sobre a população de CTT de CME...

Mucoepidermoid carcinoma (MEC) is the most common malignant salivary tumor compromising about 30% of all salivary malignances. Managing MEC patients remain challenging especially due to the heterogeneous response of tumor cells to available therapy. For this reason clinical outcome remains unpredictable. Current treatment of MEC encompasses surgical resection with eventual adjuvant radiotherapy, which frequently leads to functional and aesthetic complications. The use of chemotherapy is often reserved for recurrent and metastatic tumors. Administration of single-agent or combination therapy has showed activity, however overall response rates are unsatisfactory and of short duration. Emerging evidences suggest that the modest response of tumor cells to therapy resulting in high recurrence rates and poor survival, are associated with the presence of cancer stem cells (CSC). Quiescence of CSC is achieved by the reduced levels of transcription in a process that requires tight folding of DNA driven by core histone proteins. Changes in DNA folding are responsible for different cellular phenotypes mediated by a cell type-specific chromatin organization. Of interest, we also found that acetylation of HNSCC tumor histones driven by histone deacetylase (HDAC) inhibitors abrogate tumor resistance to chemotherapy. We investigate the effects of HDACi and cisplatin in the population of CSCs of MEC...

Effects of SAHA combined with PTX on survival and apoptosis of human paclitaxel-resistant ovarian cancer cells / 军事医学

Objective To evaluate the effect of SAHA or/and PTX on survival and apoptosis of human paclitaxel-resist-ant ovarian cancer OC3/P cells, and explore whether the combination of two drugs has a synergistic effect .Methods The morphology of OC3/P cells in different drug-groups was observed by inverted microscope .Cell viability was evaluated by MTT assay.The apoptosis rate was analyzed by Annexin V-FITC/PI assay.Results The morphology change of OC 3/P cells treated with different drug was observed by inverted microscope , and the change in combination group was more signif-icant than one drug alone group .The result of cell survival measured by MTT assay showed that inhibition rate of combina -tion group was more higher than one drug alone group (P<0.05).The analysis of factorial design and gold formula method all proved that the two drugs had synergy .Further the result of flow cytometry showed that apoptosis rate in combination group was significantly higher than SAHA or PTX alone group (P<0.05).Conclusion SAHA and PTX can inhibit the survival and induce apoptosis of OC 3/P cells, and two drugs have synergistic antitumor effects .

RPE Cell Apoptosis by the Combination Treatment of SAHA and Lactacystin

PURPOSE: To establish a new therapeutic strategy for proliferative vitreoretinopathhy (PVR), we examined the effect of combined treatment with HDAC inhibitor SAHA and proteasome inhibitor lactacystin in human retinal pigment epithelial (RPE) cells, ARPE-19. METHODS: Viability was determined by trypan blue exclusion assay. Mitochondrial membrane potential (MMP) was measured by flow cytometry. Proteasome activity was measured by fluorophotometry. The expression and degradation of apoptosis-related proteins were assesssed by Western blotting. Subcellular location of apoptosis-related factors was monitored by confocal miscroscopy. RESULTS: A single treatment with 5 micro M SAHA or 10 micro M lactacystin did not reduce cell viability. However, combination treatment with 5 micro M SAHA and 10 micro M lactacystin substantially reduced the viability, because the mixture induced the reduction of MMP and nuclear condensation or fragmentation. Moreover, the combination treatment triggered the activation of caspase-3 and the production of PARP cleavage products. These data indicate that the combination treatment efficiently induces apoptosis in ARPE-19 cells. However, co-treatment of SAHA did not augment the proteasome inhibitory activity of lactacystin, nor did co-treatment of lactacystin augment acetylation of histones. It is notable that while p53 and CAD were observed in the mitochondria of cells treated with SAHA, they were translocated into the nucleus after the combination treatment. CONCLUSIONS: These results suggest that the combination treatment of SAHA and lactacystin effectively induced apoptosis in ARPE-19 cells. Further work is warranted to develop this combination therapy as a novel therapeutic strategy for PVR.

A Case of SAHA Syndrome / 대한피부과학회지

The SAHA syndrome is an acronym which stands for seborrhea, acne, hirsutism and androgenic alopecia. The SAHA syndrome generally occurs in young to middle-aged women and may be caused by elevated blood levels of androgens or increased androgen-driven peripheral response with normal circulating androgen levels. In SAHA syndrome, careful diagnostic and clinical evaluation is necessary in order to identify the cause of peripheral hyperandrogenism, and to exclude androgen-producing tumors. SAHA can be classified into 5 subtypes: familial, ovarian, adrenal, hyperprolactinemic SAHA and HAIRAN (hyperandrogenism, insulin resistance, acanthosis nigricans) syndrome. Among them, ovarian SAHA syndrome is associated with polycystic ovarian syndrome. We report a case of ovarian SAHA syndrome in 15-year-old girl who showed seborrea, acne, hirsutism and androgenic alopecia associated with polycystic ovarian syndrome.