Journal: PLoS ONE

Article Title: RNA Binding Protein CUGBP2/CELF2 Mediates Curcumin-Induced Mitotic Catastrophe of Pancreatic Cancer Cells

doi: 10.1371/journal.pone.0016958

Figure Lengend Snippet: (A) Nude mice carrying Pan02 cell tumor xenografts in the flanks were administered curcumin intraperitoneally for 3 weeks. There was a significant reduction in tumor size from curcumin-treated animals when compared to untreated controls (*p<0.05). (B) Curcumin treatment resulted in significantly lower tumor weight when compared to controls (*p<0.05). (C) Tumor sections were stained for CD31, an endothelial cell specific surface marker and with hematoxylin and eosin (H&E). A representative figure is presented showing significant reduction in microvessels. (D) The number of microvessels in the tumor tissues were counted in 12 high power fields and averaged. Data shows that microvessel density was significantly reduced in the xenografts of curcumin treated animals (*p<0.05).

Article Snippet: AsPC-1, MiaPaCa-2, Panc-1, BxPC-3 human and Pan02 mouse pancreatic cancer cells (all from American Type Culture Collection, Manassas, VA) were grown in RPMI 1640 containing 10% heat inactivated fetal bovine serum (Sigma Chemical Co, St. Louis, MO) and 1% antibiotic-antimycotic solution (Mediatech Inc, Herndon, VA) at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Staining, Marker

Journal: PLoS ONE

Article Title: RNA Binding Protein CUGBP2/CELF2 Mediates Curcumin-Induced Mitotic Catastrophe of Pancreatic Cancer Cells

doi: 10.1371/journal.pone.0016958

Figure Lengend Snippet: (A) Curcumin inhibits proliferation of pancreatic cancer cells. MiaPaCa-2, Pan02, AsPC-1 and BxPC-3 cells were incubated with curcumin (1–50 µM) for up to 72 h. Cell proliferation was analyzed using hexosaminidase enzyme activity. Curcumin treatment resulted in a significant dose-and time-dependent decrease in cell proliferation in all the cells when compared with untreated controls. (B) Curcumin inhibits colony formation. MiaPaCa-2 and Pan02 cells were incubated with 30 µM of curcumin for 24 h and colonies were allowed to form by incubating in regular media containing 10% FBS for an additional 10 d. Treatment with curcumin inhibited colony formation. A representative of three independent experiments is shown. (C) Expression of cyclin D1 is suppressed at 24 h. RNA from MiaPaCa-2 and Pan02 cells incubated with 30 µM curcumin were subjected to Real time PCR for cyclin D1 mRNA expression. There was significant suppression of the mRNA at 24 h but not at 12 h (*p<0.05). (D) Lysates from MiaPaCa-2 or Pan02 incubated with 30 µM curcumin were analyzed by western blotting for cyclin D1 expression levels. Curcumin treatment inhibits cyclin D1 mRNA and protein expression.

Article Snippet: AsPC-1, MiaPaCa-2, Panc-1, BxPC-3 human and Pan02 mouse pancreatic cancer cells (all from American Type Culture Collection, Manassas, VA) were grown in RPMI 1640 containing 10% heat inactivated fetal bovine serum (Sigma Chemical Co, St. Louis, MO) and 1% antibiotic-antimycotic solution (Mediatech Inc, Herndon, VA) at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Incubation, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: RNA Binding Protein CUGBP2/CELF2 Mediates Curcumin-Induced Mitotic Catastrophe of Pancreatic Cancer Cells

doi: 10.1371/journal.pone.0016958

Figure Lengend Snippet: A) Cell cycle profiles of MiaPaCa-2 cells treated with curcumin for 12 h and 24 h and were determined by flow cytometry using propidium iodide staining for DNA content. Curcumin treatment significantly increased cells in the S and G 2 M phase of cell cycle within 12 h. (B) Curcumin treatment induces apoptosis. MiaPaCa-2 and Pan02 cells incubated with 30 µM of curcumin were analyzed for apoptosis by caspase 3/7 activation. Curcumin treatment increased the number of cells undergoing apoptosis compared to untreated controls (*p<0.05). (Inset) Curcumin induces caspase 3, an apoptosis mediator. Lysates from MiaPaCa-2 or Pan02 cells incubated with 30µM curcumin were analyzed by western blotting for caspase 3 protein. Curcumin treated cells shows cleaved (activated) caspase 3 while untreated cells have no cleaved caspase-3. (C) Curcumin treatment increased the levels of cyclin B1, Cdc-2 and phosphorylated checkpoint kinase Chk2. Lysates from curcumin treated MiaPaCa-2 cells (left) and Pan02 tumor xenografts (right) were analyzed by western blotting for phospho Chk2, and cyclin B1 and Cdc-2 protein expression levels. Curcumin treatment phosphorylates checkpoint kinases and increased levels of cyclin B1 and Cdc-2 in both MiaPaCa-2 cells and Pan02 tumor xenografts. (D) Curcumin treatment results in nuclear localization of cyclin B1 and Cdc-2. Immunocytochemistry of curcumin treated with MiaPaCa-2 cells both the cyclinB1 and Cdc-2 protein levels were predominately in the nucleus compared than untreated cells. Phosphorylation of Chk2, coupled with increased expression and nuclear translocation of cyclin B1 and Cdc-2 demonstrates mitotic progression of cells.

Article Snippet: AsPC-1, MiaPaCa-2, Panc-1, BxPC-3 human and Pan02 mouse pancreatic cancer cells (all from American Type Culture Collection, Manassas, VA) were grown in RPMI 1640 containing 10% heat inactivated fetal bovine serum (Sigma Chemical Co, St. Louis, MO) and 1% antibiotic-antimycotic solution (Mediatech Inc, Herndon, VA) at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Flow Cytometry, Staining, Incubation, Activation Assay, Western Blot, Expressing, Immunocytochemistry, Translocation Assay

Journal: PLoS ONE

Article Title: RNA Binding Protein CUGBP2/CELF2 Mediates Curcumin-Induced Mitotic Catastrophe of Pancreatic Cancer Cells

doi: 10.1371/journal.pone.0016958

Figure Lengend Snippet: (A) Total RNA from Pan02 tumor xenografts were subjected to Real time PCR. Curcumin treatment resulted in reduced COX-2, VEGF and cyclin D1 mRNA levels when compared to controls. On the other hand, CUGBP2 and TIA-1 mRNA expression was increased (*p<0.05). Data is an average from xenografts in 5 mice. (B) Western blot analysis demonstrated that tissue lysates from the curcumin treated animals have significantly lower levels of COX-2, VEGF, and cyclin D1 proteins but increased levels of CUGBP2 and TIA-1 proteins. (C) Immunohistochemistry demonstrates that curcumin treatment significantly reduced the expression of COX-2 and VEGF. (D) Immunohistochemistry demonstrates increased expression of CUGBP2 and TIA-1 in the tumor xenografts.

Article Snippet: AsPC-1, MiaPaCa-2, Panc-1, BxPC-3 human and Pan02 mouse pancreatic cancer cells (all from American Type Culture Collection, Manassas, VA) were grown in RPMI 1640 containing 10% heat inactivated fetal bovine serum (Sigma Chemical Co, St. Louis, MO) and 1% antibiotic-antimycotic solution (Mediatech Inc, Herndon, VA) at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunohistochemistry

Journal: Cell & Bioscience

Article Title: GPR116 receptor regulates the antitumor function of NK cells via Gαq/HIF1α/NF-κB signaling pathway as a potential immune checkpoint

doi: 10.1186/s13578-023-01005-7

Figure Lengend Snippet: Deficiency of GPR116 receptor enhances the antitumor effect of NK cells in vivo. Subcutaneous tumor models were established with PAN02 cells and these mice were injected with PBS, WT-NK and GPR116 −/− -NK through tail vein, respectively. A The photo of PAAD tumor sizes (n = 4). B The PAAD tumor growth curves and the end-point tumor sizes were represented (n = 4). C The weight of PAAD tumor (n = 4). D The proportion of CD4 + and CD8 + T cells in PAAD tumor. E The infiltration of CD3 − NK1.1 + cells in PAAD tumor. F The expression of GzmB in tumor-infiltrating CD3 − NK1.1 + cells. G The expression of IFNγ in tumor-infiltrating CD3 − NK1.1 + cells. Data are represented as the mean ± standard error of the mean (SEM). *P < 0.05, **P < 0.01, ***P < 0.001 by an unpaired Student’s t-test

Article Snippet: Mouse pancreatic carcinoma cell line PAN02 and human pancreatic carcinoma cell line PANC28, PANC1, SW1990 were purchased from the American Type Culture Collection and cultured in DMEM medium contained 10% FBS (Gibco, Gaithersburg, MD, USA) at 37 °C in an atmosphere of 5% CO 2 .

Techniques: In Vivo, Injection, Expressing

Journal: Oncoimmunology

Article Title: EP4 Antagonism by E7046 diminishes Myeloid immunosuppression and synergizes with Treg-reducing IL-2-Diphtheria toxin fusion protein in restoring anti-tumor immunity

doi: 10.1080/2162402X.2017.1338239

Figure Lengend Snippet: In vivo genetic or pharmacological blockade of EP4 results in anti-tumor activity in both a (T)cell and myeloid cell-dependent manner. A. Delayed tumor growth in EP4 −/− mice. Growth of s.c. inoculated LLC lung cancer in mice that expressed wild type (EP 4 flox/flox ), or homozygous (Cre-ER x EP 4 flox/flox ) deletion in EP4. *P = 0.003, Student's t test; n = 9–10. B. Representative results of EP4 antagonism by E7046 effect on subcutaneous Sal/N fibrosarcoma tumors (n = 8). By the end of treatment, 2 of 8 mice were tumor-free. C. Representative graph of E7046 activity on Pan02 mouse pancreatic cancer s.c. model (n = 5). D. Representative graph of E7046 activity on EMT6 mouse breast cancer orthotopic model (n = 7). E. Dose-dependent E7046 activity against breast cancer 4T1 s.c. model (n = 10). F. CD8 + T cell-dependent anti-tumor activity of E7046. CD8 + T or CD4 + T cells were depleted in CT26 tumor-bearing mice by i.p. administration of anti-CD8 or anti-CD4 antibodies (n = 11). G. Lack of anti-tumor activity of E7046 in CT26 tumor-bearing nude mice. H. Effect of myeloid cells in the anti-tumor activity of E7046. Comparable anti-tumor activities were detected in macrophage-depleted CT26 tumor-bearing mice (treated with clodronate-containing liposomes) as with mice treated with E7046. E7046 was orally administered daily at a dose of 150 mg/kg, while antibodies and liposomes were administered i.p. as shown in the graphs. * P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; 2- tailed student's t-test.

Article Snippet: Mouse cell lines CT26, LL2, 4T1, EMT6, SAI/N, and were obtained from ATCC, mouse cell line Pan02 was obtained from the DCTD-NCI (Frederick, MD) and mouse cell line H22 from GenScript (Piscataway, NJ).

Techniques: In Vivo, Activity Assay

Journal: BMC Cancer

Article Title: The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer

doi: 10.1186/1471-2407-8-352

Figure Lengend Snippet: Tumor size is reduced after CT-322 treatment in a syngeneic model of pancreatic cancer . Orthotopic murine Pan02 tumors were established in C57BL/6J wild-type mice. Treatment twice weekly i.p. injections of CT-322 (30 mg/kg), gemcitabine (3.5 mg/animal/injection) (GEM), or CT-322 (CT) + GEM was started 7 days after tumor cell injection and continued for 6 weeks, at which point animals were sacrificed. Mean (+/- SEM) tumor weight at the time of sacrifice is displayed. The combination treatment of CT-322 and GEM significantly inhibited tumor growth (control 0.44 ± 0.11 g, 0.153 ± 0.01 g; n = 8/group except CT+GEM n = 10; p < 0.05).

Article Snippet: The human pancreatic carcinoma cell line MiaPaCa-2 (ATCC, Manassas, VA) and the murine cell line Pan02 also known as Panc02 (NCI, Frederick, MD) were maintained at 37°C in a mixture of 5% CO 2 and 95% air in Dulbecco's minimal essential medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 10% fetal calf serum (Gimini Bio-Products, Woodland, CA).

Techniques: Injection

Journal: BMC Cancer

Article Title: The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer

doi: 10.1186/1471-2407-8-352

Figure Lengend Snippet: Effect of therapy on metastatic burden in mice bearing Pan02 tumors

Article Snippet: The human pancreatic carcinoma cell line MiaPaCa-2 (ATCC, Manassas, VA) and the murine cell line Pan02 also known as Panc02 (NCI, Frederick, MD) were maintained at 37°C in a mixture of 5% CO 2 and 95% air in Dulbecco's minimal essential medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 10% fetal calf serum (Gimini Bio-Products, Woodland, CA).

Techniques:

Journal: BMC Cancer

Article Title: The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer

doi: 10.1186/1471-2407-8-352

Figure Lengend Snippet: VEGFR2 inhibition with CT-322 reduces microvessel density in a syngeneic model of pancreatic cancer . Frozen sections of tumors from mice treated with vehicle alone (Control), CT-322, gemcitabine (GEM), or the combination of CT-322 + gemcitabine were evaluated for microvessel density by immunohistochemistry using MECA-32. Representative images of MECA-32 immunohistochemistry are displayed along with the mean (+/- SEM) number of blood vessels/field (total magnification, 200×). Microvessel density in Pan02 tumors from mice treated with CT-322 monotherapy was reduced compared to tumors from control-treated animals (control, 70.0 ± 3.6; CT-322, 52.6 ± 2.3; 3 tumors per group, at least 5 fields per tumor; *, p < 0.001 vs. control). Two representative pictures from each group are shown.

Article Snippet: The human pancreatic carcinoma cell line MiaPaCa-2 (ATCC, Manassas, VA) and the murine cell line Pan02 also known as Panc02 (NCI, Frederick, MD) were maintained at 37°C in a mixture of 5% CO 2 and 95% air in Dulbecco's minimal essential medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 10% fetal calf serum (Gimini Bio-Products, Woodland, CA).

Techniques: Inhibition, Immunohistochemistry