Synergistic effect of docetaxel combined with a novel multi-target inhibitor CUDC-101 on inhibiting human prostate cancer.

Histone deacetylases (HDACs) are commonly overexpressed in several types of human cancers, including prostate cancer (PCa). Histone deacetylase inhibitors (HDACis) are emerging as promising tools for cancer therapy. However, there is still a need to understand their anti-tumor effects and the mechanisms underlying their action. In our study, we investigated the effects of co-treatment with CUDC-101 and docetaxel (DTX) on cell growth, clonogenicity, invasion and migration of PCa cells both in vitro, and in a xenograft mouse model. We found that the combination of CUDC-101 and DTX significantly reduced tumor growth, as evidenced by lower tumor weight and volumes. Moreover, apoptotic cell death was increased in the combination group compared to either drug alone or control. Mechanistically, we observed that the combined treatment of CUDC-101 with DTX suppressed the progression of PCa cell lines through the AKT and ERK1/2 signaling pathways. Additionally, this combination treatment reversed EMT by modulating the expression of key markers such as E-cadherin, vimentin, Snail and MMP-9. To conclude, these results demonstrated that the combination of CUDC-101 with DTX had a synergistic and significantly improved anti-carcinogenic effect. This combination may serve as a potential strategy for clinical treatment and prognosis improvement in PCa.

Comparison of magnetic susceptibility probes relevantly used in soil contamination applications.

Magnetic susceptibility (MS) technology can achieve the efficient rough measurement, mapping, and pollution assessment of soil heavy metal concentrations in topsoil due to atmospheric dust contamination. However, previous studies of commonly used MS field probes (MS2D, MS2F, and MS2K) have not dealt with the range of magnetic signal detection and the attenuation characteristics of the signal with respect to distance. In this study, the vertical and horizontal measurement ranges of the MS2D, MS2F, and MS2K probes were explored through laboratory and field experiments, and the intensity of their magnetic signals was further compared and analyzed in the field. The results showed that the magnetic signal intensity of the three probes decreased exponentially with distance. The penetration depths of the MS2D, MS2F, and MS2K probes were 8.5, 2.4, and 3.0 cm, respectively, and the horizontal detection boundary lengths of their magnetic signals were 32, 8, and 6.8 cm, respectively. In the field surface soil MS detection, the magnetic measurement signals of the MS2F and MS2K probes showed a weak linear correlation with the MS2D probe (R2 of 0.43 and 0.50, respectively), while the MS2F and MS2K probes had a significantly better correlation (R2 = 0.68) with each other. In general, the MS2D probe and MS2K probe correlation had a slope close to unity, meaning MS2K probes had good mutual substitution. Furthermore, results of this study improve the effectiveness of the MS evaluation of heavy metal pollution in urban topsoil.

The risk characteristics of heavy metals in urban soil of typical developed cities in China.

With the rapid development of China's industrial economy, heavy metals continue to accumulate in the environment, which has created serious threats for the ecological environment and human health. This study collected 50 surface soil samples in Nanjing, a typical developed city in China, and the contents of Al, Ca, Fe, Mg, Mn, Ni, Ti, Cd, Cr, Cu, Pb, and Zn in the samples were determined. Combined with the ecological risk index and the health risk assessment model, the risk of soil heavy metals in Nanjing was comprehensively evaluated. The results show that the variation coefficients of Pb and Cu are distinctly large, and these elements are all slightly polluting. Children are at a high risk of exposure in various ways, among which Pb and Cu elements have a high risk of causing non-carcinogenic issues. The results of the correlation analysis showed that the content changes of Pb, Zn, and Cu had extremely significant correlations, indicating that they may have the same source. The results of the principal component analysis showed that industrial sources in Nanjing contributed the most heavy metals, reaching 34.4%. The second largest source was from parent material and fertilizer, which contributed 32.3% and 19.6%, respectively. The sources with the lowest contributions were from weathering and deposition, which reached 13.7%. The results of this study will provide guidance and reference for risk-source analysis, early warning, and management of soil heavy metals in developed cities.

CUDC-101 enhances the chemosensitivity of gemcitabine-treated lymphoma cells.

BACKGROUND: The metastasis and recurrence of Non-Hodgkin's lymphoma (NHL) is a major cause of morbidity and mortality. Recent work suggests that drugs capable of targeting epigenetic regulatory mechanisms may be well suited to the treatment of such disease progression. METHODS: This study was thus designed to evaluate the ability of the novel histone deacetylase (HDAC) inhibitor CUDC-101 to synergize with gemcitabine in order to kill human HUT78 and Pfeiffer NHL cells. To that end, we analyzed the viability of these NHL cells via CCK-8 assay, while the incidence of apoptosis among treated cells was evaluated via Annexin V-FITC/PI staining and by the Western blotting-mediated evaluation of proteins associate with apoptosis and related signaling pathways. RESULTS: We found that CUDC-101 and gemcitabine interacted synergistically to reduce NHL cell viability and to induce the apoptotic death of these cells via the EGFR/ PI3K/Akt and Erk pathways, which were regulated by HDAC signaling pathways. CONCLUSION: Together, our results highlight the anti-cancer properties of CUDC-101 alone or in combination with gemcitabine as an approach to inducing the apoptotic death of lymphoma cells in vitro, while also offering insight into the underlying molecular mechanisms governing this activity.

Bone marrow mesenchymal stem cell derived exosomes miR-21-5p promotes proliferation, migration and invasion of prostate cancer PC-3 cell by downregulating PHLPP2 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of exosome originated from bone marrow mesenchymal stem cell (BMSCs) on proliferation, migration and invasion of prostate cancer PC-3 cell and its mechanism. Methods: qPCR was used to detect the expression level of miR-21-5p in prostate cancer cell lines. The morphology of exosomes isolated from BMSCs was observed with an electron microscope. Western blotting was used to detect the expressions of exosome surface markers and the epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin and Vimentin). Dual luciferase reporter gene experiment was used to detect the targeted regulation relationship between miR-21-5p and PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2). PC-3 cells were co-cultured with 10 μl BMSCs exosomes suspension (Exo group), transfected with sh-PHLPP2 or antagomiR, then CCK-8 and Transwell experiments were used to detect changesinproliferation,migrationandinvasionofPC-3cell.Results: miR-21-5p was highly expressed in prostate cancer PC-3 cell line. The exosomes in the supernatant of BMSCs culture fluid were successfully isolated, and the typical vesicle-like structures of exosomes were observed under transmission electron microscope. Exosomes expressed specific proteins such as CD9, CD63 and CD81. In the Exo group, the proliferation, invasion, migration, as well as the expressions of N-cadherin, Vimentin and miR-21-5p in PC-3 cells were significantly higher than those in the control group (all P<0.05). PHLPP2 is a target gene of miR-21-5p. Compared with the control group, the expression of PHLPP2 in PC-3 cells of Exo group and sh-PHLPP2 group was significantly reduced (0.66±0.09, 0.42±0.05 vs 1.09±0.08, all P<0.01); cell viability, invasion and migration were significantly improved (all P<0.01); and E-cadherin expression level was significantly reduced while N-cadherin and Vimentin expressions were significantly increased (both P<0.05). Conclusion: miR-21-5p is highly expressed in prostate cancer PC-3 cell line. BMSC exosome miR-21-5p can increase the proliferation, migration and invasion ability of PC-3 cells through targeted down-regulation of PHLPP2.

HDAC inhibitors, trichostatin A and valproic acid, increase E­cadherin and vimentin expression but inhibit migration and invasion of cholangiocarcinoma cells.

The effects of histone deacetylase (HDAC) inhibitors on epithelial-mesenchymal transition (EMT) differ in various cancers. E­cadherin is a cell­to­cell adhesion protein, whereas accumulation of vimentin is related to the development of the spindle shape of the mesenchymal cell phenotype. We investigated the EMT phenotypes of human cholangiocellular carcinoma HuCC­T1, JCK and SNU­1079 cell lines. To this end, we measured the expression of E­cadherin or zonula occludens (ZO)­1 and vimentin, epithelial and mesenchymal cell markers, respectively, using real­time reverse transcription­polymerase chain reaction, western blotting, and immunofluorescence microscopy following treatment with trichostatin A (TSA, 200 nM) or valproic acid (VPA, 0.5 mM) with or without gemcitabine (GEM, 50 nM) for 24 h. In addition, we performed cell morphology, migration, and invasion assays. HuCC­T1 cells changed from spindle­ to rectangular­shaped after co­treatment with GEM and TSA or VPA. Furthermore, cells co­treated with GEM and TSA or VPA exhibited protein levels of E­cadherin or ZO­1 that were higher than those in cells treated with GEM alone, indicating stronger inhibition of EMT. However, vimentin expression was also increased. Confocal microscopy revealed enhanced expression of E­cadherin or ZO­1 and vimentin in all three cell lines. Migration and invasion were inhibited in HuCC­T1 cells co­treated with GEM and TSA or VPA, compared to those treated with GEM alone. In conclusion, co­treatment of cholangiocarcinoma cells with TSA or VPA and GEM suppressed EMT with tolerable cytotoxicity. However, the HDAC inhibitors augmented both E­cadherin and vimentin expression and their effects varied in different cholangiocarcinoma cell lines. Therefore, the clinical use of HDAC inhibitors in biliary cancer should be considered cautiously.

Antitumor activity of the novel HDAC inhibitor CUDC-101 combined with gemcitabine in pancreatic cancer.

Histone deacetylase (HDAC) is overexpressed in multiple cancers including pancreatic cancer (PC). However, the effects of histone deacetylase inhibitor (HDACi) on apoptosis and epithelial-mesenchymal transition (EMT) differ in various cancers. In this study, we aimed to investigate the anti-tumor effects of a novel multitargets HDACi, CUDC-101, combined with gemcitabine in PC cell lines. In vitro, we found that Co-treatment with CUDC-101 and gemcitabine results in greater levels of apoptosis and significantly inhibited cell proliferation on PC cells. In addition, CUDC-101 enhanced gemcitabine-induced apoptosis via inhibited PI3K/Akt/mTOR and Erk pathway activation, as indicated by the phosphorylation status of Akt, 4EBP1, S6 and Erk. We also found that co-treatment with gemcitabine and CUDC-101 not only synergistically suppressed ability of PC cell migration and invasion, but also synergistically inhibited EMT signaling pathway through modulation of cadherin, vimentin and transcription factors Snail, Slug and MMP-9. In vivo, the co-treatment group showed a significant anti-tumor function in the growth of xenograft tumors. Overall, combination of CUDC-101 and gemcitabine significantly increased anti-tumor activities compared with single drug alone, thus supporting a further evaluation of combination treatment for PC. Accordingly, it provides a rationale to investigate the combination of gemcitabine and CUDC-101 as a potential therapeutic strategy for PC.

VPApromotesEMTincoloncarcinomacellsvianucleartranslocationofE-cadherin / 中国肿瘤生物治疗杂志

@# Objective: To explore the effect of histone deacetylase (HDAC) inhibitor valproic acid (VPA) on the epithelial-mesenchymal transition (EMT) of colon cancer. Methods: With four colon carcinoma cell lines (DLD-1, HCT116, SW480 and HT29) as study subjects, the effect of different concentrations of VPA(0.5，5 mmol/L) on cell proliferation was detected by MTT assay. The expression level of EMT-related proteins (E-cadherin and vimentin) was detected by Western blotting; Phenotypic changes of E-cadherin and vimentin were detected by immunofluorescence staining; Cell migration and invasion ability was detected by wound healing and Transwell invasion assay, respectively. Results:After treated with different concentrations of VPAfor 48 h, low concentration of VPAmerely exerted any effect on the cell proliferation rate of four colon cancer cell lines, and thus was chosen as the experiment concentration; The results of Western blotting showed that the expression of E-cadherin was reduced (P<0.05) and vimentin was increased (P<0.05) in colon carcinoma cells by VPAtreatment (0.5 mmol/L); Immunofluorescence staining revealed membranous attenuation or nuclear translocation of E-cadherin and enhanced expression of vimentin after VPA treatment (0.5 mmol/L), and these responses occurred after 6 h and sustained until 24 h; Wound healing and Transwell invasion assay showed increased migration and invasion ability following VPA treatment (0.5 mmol/L). Conclusion: Low concentration VPA could induce the development of EMT in colon cancer cells by nuclear translocation of E-cadherin, and obviously enhance the migration and invasion ability of colon cancer cells; Thus, HDAC inhibitors, as a new type anti-cancer option, shall be carefully considered before their application in colon cancer.

Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma.

Nicotinamide adenine dinucleotide phosphate: quinone oxidoreductase 1 (NQO1) protects cells from oxidative damage. NQO1 has been reported to be upregulated in numerous solid tumors, suggesting a role in carcinogenesis and tumor progression. The present study attempted to investigate the clinical prognostic significance of NQO1 overexpression in pancreatic ductal adenocarcinoma (PDAC). A total of 181 tissue specimens were studied, including 126 PDAC and 55 normal pancreas specimens, which were selected for immunohistochemical staining of NQO1 protein. Immunofluorescence staining was additionally performed to identify the subcellular localization of NQO1 protein in pancreatic cancer BxPC-3 cells. The association between NQO1 overexpression and the clinical features of PDAC were evaluated by χ2 and Fisher's exact test. Overall survival of PDAC patients was calculated using Kaplan-Meier analysis. Univariate and multivariate analyses were performed using the Cox proportional hazards regression model. The NQO1 protein was mainly located in the cytoplasm and nucleus of BxPC-3 cells. The strongly positive rate of NQO1 expression in PDAC (65.9%, 83/126) was increased compared with that in normal pancreatic tissues (10.9%, 6/55). The positive rate of NQO1 protein was associated with grading, lymph node stage and tumor-node-metastasis (TNM) stage. Additionally, multivariate analysis suggested that NQO1 was a significant independent prognostic factor along with TNM stage in PDAC. In conclusion, high expression of NQO1 appears to be associated with PDAC progression, and may be an independent prognostic biomarker in PDAC.

The clinicopathological significance of Mortalin overexpression in invasive ductal carcinoma of breast.

BACKGROUND: Mortalin/GRP75 is a ubiquitous mitochondrial chaperone which related to the cytosolic heat shock protein 70 (HSP70), and plays a role in carcinogenesis. This study aims to investigate the Mortalin expression in breast cancer and its correlation with the outcome of the patients with breast cancer. METHODS: A total of 155 invasive ductal carcinoma of breast patients with strict follow-up, 52 ductal carcinoma in situ (DCIS) and 45 adjacent non-tumor breast tissues were selected for immunohistochemical (IHC) staining of Mortalin protein. The localization of Mortalin protein was detected in MDA-MB231 breast cancer cells using immunofluorescence (IF) staining. The correlations between overexpression of Mortalin and the clinical features of patients with breast cancer were evaluated using chi-square test and Fisher's exact tests. The survival rates were calculated by the Kaplan-Meier method, and the relationship between prognostic factors and patient survival was also analyzed by the Cox proportional hazard models. RESULTS: Mortalin protein showed a mainly cytoplasmic staining pattern in breast cancers by using IHC staining in paraffin embedded breast cancer tissues and IF staining in MDA-MB231 breast cancer cells. The strongly positive rate of Mortalin protein was 63.9% (99/155) in invasive ductal carcinoma of breast and was significantly higher than in DCIS 34.6% (18/52) and adjacent non-tumor tissues 15.6% (7/45). Overexpression of Mortalin was closely correlated with histological grade, clinical stage, lymph node metastasis, lower disease free survival (DFS) and overall survival (OS) rates of patients with breast cancer. Moreover, multivariate analysis suggested that Mortalin emerged as a significant independent prognostic factor along with clinical stage and Her2 expression status in patients with breast cancer. CONCLUSIONS: Mortalin is upregulated in breast cancer, and may be a useful poor prognostic biomarker as well as a potential therapeutic target for patients with breast cancer.

HDAC inhibitors induce epithelial-mesenchymal transition in colon carcinoma cells.

The effects of histone deacetylase (HDAC) inhibitors on epithelial-mesenchymal transition (EMT) differ in various types of cancers. We investigated the EMT phenotype in four colon cancer cell lines when challenged with HDAC inhibitors trichostatin A (TSA) and valproic acid (VPA) with or without transforming growth factor-ß1 (TGF-ß1) treatment. Four colon cancer cell lines with different phenotypes in regards to tumorigenicity, microsatellite stability and DNA mutation were used. EMT phenotypes were assessed by the expression of E-cadherin and vimentin using western blot analysis, immunofluorescence, quantitative real-time RT-PCR following treatment with TSA (100 or 200 nM) or VPA (0.5 mM) with or without TGF-ß1 (5 ng/ml) for 24 h. Biological EMT phenotypes were also evaluated by cell morphology, migration and invasion assays. TSA or VPA induced mesenchymal features in the colon carcinoma cells by a decrease in E-cadherin and an increase in vimentin expression at the mRNA and protein levels. Confocal microscopy revealed membranous attenuation or nuclear translocation of E-cadherin and enhanced expression of vimentin. These responses occurred after 6 h and increased until 24 h. Colon cancer cells changed from a round or rectangular shape to a spindle shape with increased migration and invasion ability following TSA or VPA treatment. The susceptibility to EMT changes induced by TSA or VPA was comparable in microsatellite stable (SW480 and HT29) and microsatellite unstable cells (DLD1 and HCT116). TSA or VPA induced a mesenchymal phenotype in the colon carcinoma cells and these effects were augmented in the presence of TGF-ß1. HDAC inhibitors require careful caution before their application as new anticancer drugs for colon cancers.

Epithelial-mesenchymal transition-related protein expression in biliary epithelial cells associated with hepatolithiasis.

BACKGROUND AND AIM: Epithelial-mesenchymal transition (EMT) of biliary epithelial cells (BECs) plays major roles in many cholangiopathies. This study evaluated whether EMT of BECs has a role in hepatolithiasis-induced biliary fibrosis and types of BECs that are involved. METHODS: The expression of EMT-related proteins and epidermal growth factor receptor was evaluated by immunohistochemistry of liver tissues from 102 patients with hepatolithiasis, 32 patients with post-hepatitis cirrhosis, and 48 normal livers. Antibodies against E-cadherin, ß-catenin, and cytokeratin were used to identify epithelial cells and antibodies against vimentin, S100A4, podoplanin, and α-smooth muscle actin (α-SMA) were used to identify mesenchymal cells. The relationship between clinical and histological parameters and immunohistochemistry findings in BECs, and the surrounding stroma were evaluated. RESULTS: Loss of E-cadherin and acquisition of S100A4 and vimentin were observed in BECs. In all BECs, cytokeratin and ß-catenin expression were unchanged, while podoplanin and α-SMA were not expressed. Although hepatic fibrosis was more severe in post-hepatitis cirrhosis, EMT of BECs was more widespread in hepatolithiasis. In hepatolithiasis, EMT-related proteins were more highly expressed in small bile ducts than in medium or large bile ducts. Their expression was associated with the severity of biliary fibrosis and the expressions of epidermal growth factor receptor. Expression of α-SMA in fibroblasts from the portal space was closely linked to pathological changes in small bile ducts and EMT-related protein expressions in BECs. CONCLUSIONS: Proliferating cholangiocytes that form small bile ducts may contribute to cholangiopathies in hepatolithiasis through an EMT-like phenomenon or through interactions with stromal myofibroblasts.

Combined aberrant expression of E-cadherin and S100A4, but not ß-catenin is associated with disease-free survival and overall survival in colorectal cancer patients.

BACKGROUND/AIMS: Epithelial-to-mesenchymal transition (EMT) in cancers is related to metastasis, recurrence, and poor prognosis. We evaluated whether EMT-related proteins can act as prognostic biomarkers in colorectal cancer (CRC) patients. METHODS: We evaluated the expression of E-cadherin, ß-catenin, and S100A4 by immunohistochemistry (IHC) in 333 CRC tissues from the tumor center and invasive margin. Tumor budding, cell grade, tumor stage, type of tumor growth, peritumoral lymphocyte infiltration (TLI), and perineural- or lymphovascular invasion were evaluated as pathological parameters. mRNA levels of E-cadherin, N-cadherin, ß-catenin, and S100A4 from 68 specimens from the same set were analyzed by real time quantitative RT-PCR. RESULTS: Loss of E-cadherin, nuclear ß-catenin, and gain of S100A4 were higher in the invasive margin than in the tumor center. Loss of E-cadherin was associated with cell grade, macroscopic type, perineural invasion, and tumor budding, ß-catenin with microsatellite instability and tumor site, and S100A4 with growth type, macroscopic type, AJCC stage, lymphovascular invasion, and perineural invasion. The aberrant expression of E-cadherin and S100A4 not ß-catenin in the invasive margin was a significant and independent risk factor for disease-free and overall-survival by multivariate analysis, along with AJCC stage and perineural invasion. mRNA levels of ß-catenin and S100A4 were correlated with the IHC findings at the tumor invasive margin. E-cadherin and N-cadherin showed a weak inverse correlation. CONCLUSIONS: The combination of loss of E-cadherin and gain of S100A4 in the tumor invasive margin can be used to stratify patients with the same AJCC stage into different survival groups.