Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: GI 50 and treatment doses of gemcitabine and XCT790 in each cell line.

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques:

Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: A novel in vivo drug-response assay, mini-PDX, was performed and demonstrated the synergism between gemcitabine and XCT790 in PC. A, An overview of the protocol of the generation of mini-PDX pattern. B, Chest scans of the PC patient before and 10 days after surgical resection. C, The PC tissues for mini-PDX were identified as ERRα high-expressing by IHC according to our previous scoring method . D, The body weight of the animals treated with gemcitabine and/or XCT790 didn't exhibit remarkable variations compared with that of mice treated with placebo. E, The combination of gemcitabine and XCT790 exhibited synergistic effect and greater inhibitory effects on mini-PDX models than their single treatment. The data were analyzed using Student's t-tests. q score is described as bar graph and > 1.0 denotes synergism. * P < 0.05, NS: not significant in contrast to the NC group.

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques: In Vivo, Expressing

Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: Gemcitabine and XCT790 exerted synergistic cytotoxicity in PC lineage cells. A, Gemcitabine and XCT790 single treatment inhibited cell viability and proliferation of PC cell lines and HPNE in a dosage-reliant and time-reliant way. Cells were exposed to a variety of thickness of gemcitabine or XCT790 for 24, 48 or 72 hours, whose activity was detected by CCK-8 analysis. B, The combination of gemcitabine and XCT790 showed synergistic effect in PC cell lines. Gemcitabine and XCT790 were used in combination to treat PC cell lines and HPNE cells at the indicated concentrations for 48 hours. Normalized isobologram was plotted by CompuSyn Software, CI values < 1.0 demonstrated that the combinations are synergistic. C, The combination of gemcitabine and XCT790 showed synergistic inhibition of PC cell colony formation. The data were analyzed using Student's t-tests. All outcomes are expressed as average ± SD of 3 separate assays. CI value < 1 indicates synergism; q score is described as bar graph and > 1.0 denotes synergism. * P < 0.05, ** P < 0.01, *** P < 0.001 in contrast to the NC group; a and b represent to compare with the GEM or XCT group, P ˂ 0.05, respectively.

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques: Activity Assay, CCK-8 Assay, Software, Inhibition

Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: Gemcitabine and XCT790 synergistically induced G0/G1 cellular cycle retardation and apoptotic activity in PC cells. A and D, Cellular cycle analysis and apoptosis analysis were completed via flow cell technique. B and E, All q values were > 1.0, suggesting the synergistic effect of gemcitabine + XCT790 on induction of G0/G1 retardation and apoptotic activity in PC cells. C and F, Immunoblotting was applied to identify G0/G1 cellular cycle-related and apoptotic activity-related markers. The data were analyzed using Student's t-tests. The numerical results were expressed as the average ± SD of 3 separate assays. q score is described as bar graph and > 1.0 denotes synergism. * P < 0.05, ** P < 0.01, *** P < 0.001 in contrast to the NC group; a and b represent to compare with the GEM or XCT group, P ˂ 0.05, respectively.

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques: Activity Assay, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: Gemcitabine and XCT790 suppressed migration, invasion and EMT process synergistically. (A) Transwell migration analysis and (C) wound healing analysis were carried out to assess the effect of gemcitabine, XCT790 or their combination on PC cells' migratory ability. (B) Transwell invasion assay was conducted to evaluate the PC cells' invasive ability. (D) Western blot analysis was performed to examine epitheliums biomarkers (ZO-1 and E-cadherin) and mesenchyma biomarkers (N-cadherin, vimentin, snail and MMP2) after treatment with gemcitabine and/or XCT790. The data were analyzed using Student's t-tests. q score is described as bar graph and > 1.0 denotes synergism. * P < 0.05, ** P < 0.01, *** P < 0.001 in contrast to the NC group; a and b represent to compare with the GEM or XCT group, P ˂ 0.05, respectively.

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques: Migration, Transwell Invasion Assay, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: Gemcitabine and XCT790 triggered synergistic anticancer activity via inhibiting ERRα and MEK/ERK signaling pathway. A and B, The expressing levels of ERRα and MEK/ERK pathway proteins were identified via immunoblotting. C and D, The cytotoxicity induced by gemcitabine, XCT790 and their combination were significantly attenuated by ERRα overexpression and pre-treatment of ERK activator tBHQ. PC cells were incubated with gemcitabine and/or XCT790 for 2 days after stably overexpressing ERRα or pre-treated with 50 μmmol/L tBHQ for 8 hours. Cellular activity was assessed via CCK-8 analysis. The data were analyzed using Student's t-tests(*P < 0.05, **P < 0.01 and ***P < 0.001).

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques: Activity Assay, Expressing, Western Blot, Over Expression, Incubation, Stable Transfection, CCK-8 Assay

Journal: International Journal of Medical Sciences

Article Title: Gemcitabine and XCT790, an ERRα inverse agonist, display a synergistic anticancer effect in pancreatic cancer

doi: 10.7150/ijms.68404

Figure Lengend Snippet: Gemcitabine and XCT790 potently inhibited in vivo PC tumor growth. A, Images of the xenograft models and harvested tumors. B and C, Gemcitabine and XCT790 synergistically suppressed tumor growth. D, No significant change of the body weight was observed in all treated mice, suggesting that the gemcitabine and/or XCT790 were well-tolerated. E and F, Western blot analysis and IHC staining were conducted in the indicated xenograft tumors. The data were analyzed using Student's t-tests. The numerical results were expressed as the average ± SD of 3 independently performed assays. q score is described as bar graph and > 1.0 denotes synergism. * P < 0.05, ** P < 0.01, *** P < 0.001, NS: not significant in contrast to the NC group; a and b represent to compare with the GEM or XCT group, P ˂ 0.05, respectively.

Article Snippet: Gemcitabine and tBHQ were obtained from MedChemExpress, XCT790 and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.

Techniques: In Vivo, Western Blot, Immunohistochemistry

Journal: The FEBS journal

Article Title: Regulation of kynurenine biosynthesis during influenza virus infection.

doi: 10.1111/febs.13966

Figure Lengend Snippet: Fig. 8. Effect of antiviral compounds and IDO1 inhibitors on KP in IAV-infected human macrophages. A. Schematic representation showing the effect of antiviral compounds on virus entry and viral RNA transcription and replication. Two IDO inhibitors targeting IDO1 are also shown. B. Macrophages were treated with 2 μM obatoclax, 0.1 μM SNS-032, 3 μM SaliPhe, 0.1 μM JNJ872, 1 μM gemcitabine, 1 μM epacadostat or 1 μM NLG919 or remained non-treated and infected with mock or IAV (moi 3). After 24 h cell viability was measured by CTG and results were plotted. Mean ± SD, n=3. C. Cells were treated as for panel B. RNA was extracted after 8 h post infection and the expression of cellular IDO1, IFNB1 and viral M1 genes was analysed using RT-qPCRs. Mean ± SD, n=3. Statistically significant (p<0.05, T-test) differences in gene expression between virus- and mock- infected cells are indicated with asterisks. D. Cells were treated as for panel B. Media were collected after 24 h post infection, and metabolite levels were determined by LC-MS/MS. A heat map of selected metabolites is shown. Rows represent metabolites, columns represent treatments. Each cell is coloured according to the log2-transformed values of samples, expressed as fold-change relative to the average of mock controls.

Article Snippet: Obatoclax, gemcitabine, SNS032, epacadostat and NLG919 were from Selleck Chemicals, USA.

Techniques: Infection, Virus, Expressing, Gene Expression, Liquid Chromatography with Mass Spectroscopy, Transformation Assay

Journal: Biomedicines

Article Title: Expression of Claudin-9 (CLDN9) in Breast Cancer, the Clinical Significance in Connection with Its Subcoat Anchorage Proteins ZO-1 and ZO-3 and Impact on Drug Resistance

doi: 10.3390/biomedicines11123136

Figure Lengend Snippet: Implication of CLDN9 on breast cells’ response to chemotherapeutic agents over broad concentration ranges. ( A ) MDA-MB-231 cells (gemcitabine 8 μM); ( B ) MCF7 cells (gemcitabine 200 μM); ( C , D ): SKBR3 cells (gemcitabine 10 μM and cisplatin 100 μM). CT: control; KD: knockdown.

Article Snippet: A mouse monoclonal antibody to ZO-1(33-9100) was, respectively, purchased from Thermo Fisher Scientific Inc. (Thermo Fisher Scientific, Loughborough, UK). siRNA targeting human CLDN9 was obtained from Santa-Cruz Biotechnologies Inc. Chemo-drugs, including gemcitabine (GEM), docetaxel (DOC), cisplatin (CIS), methotrexate (MTX) and docetaxel were purchased from Sigma-Aldrich (Dorset, UK).

Techniques: Concentration Assay, Control, Knockdown

Journal: Biomedicines

Article Title: Expression of Claudin-9 (CLDN9) in Breast Cancer, the Clinical Significance in Connection with Its Subcoat Anchorage Proteins ZO-1 and ZO-3 and Impact on Drug Resistance

doi: 10.3390/biomedicines11123136

Figure Lengend Snippet: IC 50 of chemotherapy drugs in breast cancer cell lines with modified CLDN9 expression.

Article Snippet: A mouse monoclonal antibody to ZO-1(33-9100) was, respectively, purchased from Thermo Fisher Scientific Inc. (Thermo Fisher Scientific, Loughborough, UK). siRNA targeting human CLDN9 was obtained from Santa-Cruz Biotechnologies Inc. Chemo-drugs, including gemcitabine (GEM), docetaxel (DOC), cisplatin (CIS), methotrexate (MTX) and docetaxel were purchased from Sigma-Aldrich (Dorset, UK).

Techniques: Modification, Expressing, Control

Journal: Biomedicines

Article Title: Expression of Claudin-9 (CLDN9) in Breast Cancer, the Clinical Significance in Connection with Its Subcoat Anchorage Proteins ZO-1 and ZO-3 and Impact on Drug Resistance

doi: 10.3390/biomedicines11123136

Figure Lengend Snippet: Inhibition of Her-2 and SKBR3 cell’s response to chemotherapeutic drugs in relation with CLDN9 expression. SKBR3 wild-type cells were tested on its wild type (Her-2-positive and CLDN9-positive) and its submodel, CLDN9 knockdown (CLDN9-KD). Neratinib (NER) had a significant inhibitory effects on the growth of both models. However, following blocking of Her-2 with neratinib, the CLDN9-KD SKBR3 cells became more sensitive to Gemcitabine ( A ), Methotrexate ( B ), Docetaxel ( C ), Cisplatin ( D ) (NER plus the respective drugs in SKBR3/CLDN9-KD cells). ** p < 0.01, *** p < 0.001 vs. control cells.

Article Snippet: A mouse monoclonal antibody to ZO-1(33-9100) was, respectively, purchased from Thermo Fisher Scientific Inc. (Thermo Fisher Scientific, Loughborough, UK). siRNA targeting human CLDN9 was obtained from Santa-Cruz Biotechnologies Inc. Chemo-drugs, including gemcitabine (GEM), docetaxel (DOC), cisplatin (CIS), methotrexate (MTX) and docetaxel were purchased from Sigma-Aldrich (Dorset, UK).

Techniques: Inhibition, Expressing, Knockdown, Blocking Assay, Control

Journal: Biomedicines

Article Title: Expression of Claudin-9 (CLDN9) in Breast Cancer, the Clinical Significance in Connection with Its Subcoat Anchorage Proteins ZO-1 and ZO-3 and Impact on Drug Resistance

doi: 10.3390/biomedicines11123136

Figure Lengend Snippet: Inhibition of Her-2 and MCF7 cells’ response to chemotherapeutic drugs in relation with CLDN9 expression. MCF7 cells were tested on its wild type (Her-2 negative and CLDN9 positive) and its submodel, CLDN9 knockdown (CLDN9-KD). Neratinib (NER) had some inhibitory effects on the growth of both models. However, blocking Her-2 with neratinib did not influence cells response to Gemcitabine ( A ), Methotrexate ( B ), Docetaxel ( C ), Cisplatin ( D ) in both cell models. ns > 0.05, ** p < 0.01, *** p < 0.001 vs. control cells.

Article Snippet: A mouse monoclonal antibody to ZO-1(33-9100) was, respectively, purchased from Thermo Fisher Scientific Inc. (Thermo Fisher Scientific, Loughborough, UK). siRNA targeting human CLDN9 was obtained from Santa-Cruz Biotechnologies Inc. Chemo-drugs, including gemcitabine (GEM), docetaxel (DOC), cisplatin (CIS), methotrexate (MTX) and docetaxel were purchased from Sigma-Aldrich (Dorset, UK).

Techniques: Inhibition, Expressing, Knockdown, Blocking Assay, Control

Journal: Biomedicines

Article Title: Expression of Claudin-9 (CLDN9) in Breast Cancer, the Clinical Significance in Connection with Its Subcoat Anchorage Proteins ZO-1 and ZO-3 and Impact on Drug Resistance

doi: 10.3390/biomedicines11123136

Figure Lengend Snippet: Inhibition of Her-2 and MDA-MB-231 cells’ response to chemotherapeutic drugs in relation with CLDN9 expression. MDA-MB-231 cells were tested on its wild type (Her-2-negative and CLDN9-positive) and its submodel, CLDN9 knockdown (CLDN9-KD). Neratinib (NER) had some inhibitory effects on the growth of both models. However, blocking Her-2 with neratinib did not influence cell responses to Gemcitabine ( A ), Methotrexate ( B ), Docetaxel ( C ), Cisplatin ( D ) in both cell models. ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control cells.

Article Snippet: A mouse monoclonal antibody to ZO-1(33-9100) was, respectively, purchased from Thermo Fisher Scientific Inc. (Thermo Fisher Scientific, Loughborough, UK). siRNA targeting human CLDN9 was obtained from Santa-Cruz Biotechnologies Inc. Chemo-drugs, including gemcitabine (GEM), docetaxel (DOC), cisplatin (CIS), methotrexate (MTX) and docetaxel were purchased from Sigma-Aldrich (Dorset, UK).

Techniques: Inhibition, Expressing, Knockdown, Blocking Assay, Control

Journal: Cancer Research

Article Title: NPEPPS Is a Druggable Driver of Platinum Resistance

doi: 10.1158/0008-5472.CAN-23-1976

Figure Lengend Snippet: Project overview and synthetic lethal screen results. A, Human bladder cancer cell lines were made resistant to cisplatin, gemcitabine, or gemcitabine plus cisplatin through dose escalation. All cell lines were profiled using -omic technologies. The gemcitabine plus cisplatin-resistant cells were subjected to a pooled CRISPR screen to identify synthetic lethal gene-to-drug relationships. B, Aggregate gene set enrichment results for the synthetic lethal screen ranked by log 2 -fold change across all cell lines reveal DNA damage response and repair pathways. Each tick mark represents a gene in the associated pathway. The bars at the right are normalized enrichment scores (NES), with the FDR-corrected P values reported in the bars. C, The intersection across the CRISPR screen results identified 46 common synthetic lethal genes; all counts and gene annotations are reported in Supplementary Fig. S2. D, The percentage change in the aggregate of the sgRNAs targeting the 46 commonly synthetic lethal genes are reported across PBS or gemcitabine plus cisplatin treatment arms of the CRISPR screen. Cell lines are coded with the same colors throughout all figures.

Article Snippet: Cisplatin (Sigma) and gemcitabine hydrochloride (BOC Sciences) were both resuspended in 0.9% PBS and stored protected from light at −80°C as individual aliquots.

Techniques: CRISPR

Journal: Cancer Research

Article Title: NPEPPS Is a Druggable Driver of Platinum Resistance

doi: 10.1158/0008-5472.CAN-23-1976

Figure Lengend Snippet: Genetic inhibition of NPEPPS resensitizes GemCis-resistant cells in vitro and in vivo . A and B, KU1919-GemCis (full immunoblot reported in Supplementary Fig. S4B) or T24-GemCis cells with knockdown of NPEPPS treated with increasing doses of cisplatin or gemcitabine. A total of three technical replicates per dose (mean ± SEM). C and D, KU1919 or T24 parental cells with overexpression of NPEPPS treated with increasing doses of cisplatin or gemcitabine. A total of three technical replicates per dose (mean ± SEM). Independent experiments are reported in Supplementary Fig. S4. P values comparing IC 50 values using the sum-of-squares F test. E and F, Intracellular cisplatin levels in KU1919 and T24 cells were measured after 4 hours of 10 μmol/L cisplatin treatment using CyTOF, with the number of live cells analyzed as indicated. Group comparisons were made for triplicate experiments by normalizing intracellular cisplatin levels to the GemCis-resistant or parental cells and compared using a one-way ANOVA (*, FDR < 0.05; **, FDR < 0.01; ***, FDR < 0.001; ns, nonsignificant). G, Tumor volume (mean ± SEM) of KU1919-GemCis xenografts measured over time and across four treatment groups considering nontargeting shRNA controls (shCtrl1), shRNA targeting NPEPPS (shN39), PBS vehicle control (PBS), or gemcitabine plus cisplatin treatment (GemCis). H, Survival analysis of xenograft models with a defined endpoint of a tumor volume > 2 cm 3 . The log-rank test was applied to test significance.

Article Snippet: Cisplatin (Sigma) and gemcitabine hydrochloride (BOC Sciences) were both resuspended in 0.9% PBS and stored protected from light at −80°C as individual aliquots.

Techniques: Inhibition, In Vitro, In Vivo, Western Blot, Over Expression, shRNA