nm parp7 bps bioscience 80527  (BPS Bioscience)

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 1. MICB and ADAM15 immunohistochemical staining (SABC method, magnification, x400). (A) Negative expression of MICB in normal pancreatic tissues. (B) Negative expression of ADAM15 in normal pancre atic tissues. (C) Negative membranous expression of MICB in pancreatic cancer tissues, positive staining for MICB was only diffusely located in the stroma in TNM stage IV. (D) Positive staining for MICB was localized on the membrane of tumor cells in carcinomas with TNM stage I. (E) Negative expression of ADAM15 in pancreatic cancer tissues with TNM stage I. (F) Strong expression of ADAM15 in pancreatic cancer tissues in TNM stage IV. MICB, MHC class I polypeptide-related sequence B.

Article Snippet: MICB monoclonal antibodies (sc-80527, dilution 1:100; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse anti-ADAM15 monoclonal antibodies (sc-365752, dilution 1:100; Santa Cruz Biotechnology, Inc.) were used in this study.

Techniques: Immunohistochemical staining, Staining, Expressing, Membrane, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 2. Expression of ADAM15 and MICB in ADAM15 knockdown PANC-1 cells. Cells were treated with ADAM15 siRNA (ADAM15KD) or control siRNA (Control) for 24 h, and subjected to analysis of ADAM15 expression by (A) real-time RT-PCR and (B) western blotting. At the same time, the membrane-bound MICB and sMICB was evaluated by (C) flow cytometry and (D) ELISA. *P<0.05. MICB, MHC class I polypeptide-related sequence B.

Article Snippet: MICB monoclonal antibodies (sc-80527, dilution 1:100; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse anti-ADAM15 monoclonal antibodies (sc-365752, dilution 1:100; Santa Cruz Biotechnology, Inc.) were used in this study.

Techniques: Expressing, Knockdown, Control, Quantitative RT-PCR, Western Blot, Membrane, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 3. Changes of ADAM15 and MICB expression in PANC-1 cells after exposure to gemcitabine. (A) PANC-1 cells were treated with different concentrations of gemcitabine (dotted lines) or vehicle (DMSO; solid lines) for 24 h, and the viability of the cells was evaluated by the MTT assay. Cells were treated with 0.5 µmol/l gemcitabine or vehicle (DMSO) for 24 h and ADAM15 expression was evaluated by (B) real-time RT-PCR and (C) western blotting. The membrane-bound MICB and sMICB expression was evaluated by (D) flow cytometry and (E) ELISA. (F) mRNA levels of MICB were evaluated by real-time RT-PCR. *P<0.05. MICB, MHC class I polypeptide-related sequence B; sMICB, soluble MICB.

Article Snippet: MICB monoclonal antibodies (sc-80527, dilution 1:100; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse anti-ADAM15 monoclonal antibodies (sc-365752, dilution 1:100; Santa Cruz Biotechnology, Inc.) were used in this study.

Techniques: Expressing, MTT Assay, Quantitative RT-PCR, Western Blot, Membrane, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 4 Gemcitabine-mediated shedding of MICB is ADAM15-dependent. Cells were transfected with ADAM15 siRNA (ADAM15KD) or control siRNA (Control) for 24 h and then cultured with 0.5 µmol/l of gemcitabine or vehicle (DMSO) for 24 h. (A and B) The expression of membrane-bound MICB was evaluated by flow cytometry and (C) the statistical analysis shown as the median of geometric mean fluorescence intensity (geo MFI) of MICB is presented. (D) sMICB in the culture supernatant was evaluated by ELISA. *P<0.05. MICB, MHC class I polypeptide-related sequence B; sMICB, soluble MICB.

Article Snippet: MICB monoclonal antibodies (sc-80527, dilution 1:100; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse anti-ADAM15 monoclonal antibodies (sc-365752, dilution 1:100; Santa Cruz Biotechnology, Inc.) were used in this study.

Techniques: Transfection, Control, Cell Culture, Expressing, Membrane, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 1. MICB and ADAM15 immunohistochemical staining (SABC method, magnification, x400). (A) Negative expression of MICB in normal pancreatic tissues. (B) Negative expression of ADAM15 in normal pancre atic tissues. (C) Negative membranous expression of MICB in pancreatic cancer tissues, positive staining for MICB was only diffusely located in the stroma in TNM stage IV. (D) Positive staining for MICB was localized on the membrane of tumor cells in carcinomas with TNM stage I. (E) Negative expression of ADAM15 in pancreatic cancer tissues with TNM stage I. (F) Strong expression of ADAM15 in pancreatic cancer tissues in TNM stage IV. MICB, MHC class I polypeptide-related sequence B.

Article Snippet: For the detection of membrane-bound MICB, cells were incubated with an anti-MICB-specific antibody (sc-80527, Santa Cruz Biotechnology, Inc.) and stained with phycoerythrin (PE)-goat anti-mouse immunoglobulin (BD Biosciences, Franklin Lakes, NJ, USA) as a secondary reagent and then subjected to flow cytometry.

Techniques: Immunohistochemical staining, Staining, Expressing, Membrane, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 2. Expression of ADAM15 and MICB in ADAM15 knockdown PANC-1 cells. Cells were treated with ADAM15 siRNA (ADAM15KD) or control siRNA (Control) for 24 h, and subjected to analysis of ADAM15 expression by (A) real-time RT-PCR and (B) western blotting. At the same time, the membrane-bound MICB and sMICB was evaluated by (C) flow cytometry and (D) ELISA. *P<0.05. MICB, MHC class I polypeptide-related sequence B.

Article Snippet: For the detection of membrane-bound MICB, cells were incubated with an anti-MICB-specific antibody (sc-80527, Santa Cruz Biotechnology, Inc.) and stained with phycoerythrin (PE)-goat anti-mouse immunoglobulin (BD Biosciences, Franklin Lakes, NJ, USA) as a secondary reagent and then subjected to flow cytometry.

Techniques: Expressing, Knockdown, Control, Quantitative RT-PCR, Western Blot, Membrane, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 3. Changes of ADAM15 and MICB expression in PANC-1 cells after exposure to gemcitabine. (A) PANC-1 cells were treated with different concentrations of gemcitabine (dotted lines) or vehicle (DMSO; solid lines) for 24 h, and the viability of the cells was evaluated by the MTT assay. Cells were treated with 0.5 µmol/l gemcitabine or vehicle (DMSO) for 24 h and ADAM15 expression was evaluated by (B) real-time RT-PCR and (C) western blotting. The membrane-bound MICB and sMICB expression was evaluated by (D) flow cytometry and (E) ELISA. (F) mRNA levels of MICB were evaluated by real-time RT-PCR. *P<0.05. MICB, MHC class I polypeptide-related sequence B; sMICB, soluble MICB.

Article Snippet: For the detection of membrane-bound MICB, cells were incubated with an anti-MICB-specific antibody (sc-80527, Santa Cruz Biotechnology, Inc.) and stained with phycoerythrin (PE)-goat anti-mouse immunoglobulin (BD Biosciences, Franklin Lakes, NJ, USA) as a secondary reagent and then subjected to flow cytometry.

Techniques: Expressing, MTT Assay, Quantitative RT-PCR, Western Blot, Membrane, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Molecular medicine reports

Article Title: ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells.

doi: 10.3892/mmr.2013.1272

Figure Lengend Snippet: Figure 4 Gemcitabine-mediated shedding of MICB is ADAM15-dependent. Cells were transfected with ADAM15 siRNA (ADAM15KD) or control siRNA (Control) for 24 h and then cultured with 0.5 µmol/l of gemcitabine or vehicle (DMSO) for 24 h. (A and B) The expression of membrane-bound MICB was evaluated by flow cytometry and (C) the statistical analysis shown as the median of geometric mean fluorescence intensity (geo MFI) of MICB is presented. (D) sMICB in the culture supernatant was evaluated by ELISA. *P<0.05. MICB, MHC class I polypeptide-related sequence B; sMICB, soluble MICB.

Article Snippet: For the detection of membrane-bound MICB, cells were incubated with an anti-MICB-specific antibody (sc-80527, Santa Cruz Biotechnology, Inc.) and stained with phycoerythrin (PE)-goat anti-mouse immunoglobulin (BD Biosciences, Franklin Lakes, NJ, USA) as a secondary reagent and then subjected to flow cytometry.

Techniques: Transfection, Control, Cell Culture, Expressing, Membrane, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: BMC Cancer

Article Title: Valproic acid sensitizes pancreatic cancer cells to natural killer cell-mediated lysis by upregulating MICA and MICB via the PI3K/Akt signaling pathway

doi: 10.1186/1471-2407-14-370

Figure Lengend Snippet: VPA upregulates the expression of MICA and MICB in pancreatic cancer cells. Pancreatic cancer cells were incubated with or without 1 mM VPA for 24 h. (A) Quantitative real-time RT-PCR analysis of MICA and MICB mRNA expression. (B) Flow cytometry analysis and quantification of MICA and MICB protein expression on the surface of pancreatic cancer cells. MFI, mean fluorescence intensity. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. ** P < 0.01.

Article Snippet: Rabbit polyclonal antibodies against MICA and MICB were obtained from Santa Cruz, Santa Cruz, CA, USA.

Techniques: Expressing, Incubation, Quantitative RT-PCR, Flow Cytometry, Fluorescence

Journal: BMC Cancer

Article Title: Valproic acid sensitizes pancreatic cancer cells to natural killer cell-mediated lysis by upregulating MICA and MICB via the PI3K/Akt signaling pathway

doi: 10.1186/1471-2407-14-370

Figure Lengend Snippet: PI3K/Akt signaling is required for VPA-induced upregulation of MICA and MICB in pancreatic cancer cells. (A, B) Quantitative real-time RT-PCR analysis. The VPA-induced upregulation of MICA and MICB mRNA expression were inhibited by the HER2/HER3 inhibitor lapatinib and the PI3K inhibitor LY294002, but not by the ATM/ATR inhibitor caffeine. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. * P < 0.05; ** P < 0.01; ns P > 0.05. (C) Flow cytometry analysis. The VPA-induced upregulation of MICA and MICB protein expression on the cell surface were inhibited by the PI3K inhibitor LY294002. MFI, mean fluorescence intensity; * P < 0.05. (D) Western blotting analysis. Transfection of the PI3KCA siRNA inhibited PI3KCA protein expression at 48 h post-transfection. NC, negative control; siR1-3, PI3KCA_siR sequence 1-3; ** P < 0.01. (E) Quantitative real-time RT-PCR analysis. VPA-induced upregulation of MICA and MICB mRNA expression were attenuated in PI3KCA-knockdown cells; Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. ** P < 0.01. (F) Flow cytometric analysis. VPA-induced upregulation of MICA and MICB protein expression on the cell surface were attenuated in PI3KCA-knockdown cells. MFI, mean fluorescence intensity; * P < 0.05; ** P < 0.01. (G) LDH release assay. The VPA-induced susceptibility of cancer cells to NK cell-mediated cell lysis was reduced in PI3KCA-knockdown cells. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. ** P < 0.01 and * P < 0.05 for NC vs. siR2; ▲▲ P < 0.01 and ▲ P < 0.05 for NC vs. siR3. siR2, PI3KCA siR sequence 2; siR3, PI3KCA siR sequence 3.

Article Snippet: Rabbit polyclonal antibodies against MICA and MICB were obtained from Santa Cruz, Santa Cruz, CA, USA.

Techniques: Quantitative RT-PCR, Expressing, Flow Cytometry, Fluorescence, Western Blot, Transfection, Negative Control, Sequencing, Knockdown, Lactate Dehydrogenase Assay, Lysis

Journal: BMC Cancer

Article Title: Valproic acid sensitizes pancreatic cancer cells to natural killer cell-mediated lysis by upregulating MICA and MICB via the PI3K/Akt signaling pathway

doi: 10.1186/1471-2407-14-370

Figure Lengend Snippet: VPA sensitizes pancreatic cancer cells to NK cell-mediated lysis in vivo by upregulating the expression of MICA and MICB. (A, B) Excised xenograft tumors and growth curves for the xenografts; VPA enhanced the growth inhibitory effect of NK cells in vivo ; this effect was attenuated by the PI3K inhibitor LY294002. Data are mean ± SD of five mice per group; * P < 0.05 for NK-92 vs. NK-92 + VPA; Δ P < 0.05 for NK-92 + VPA + LY294002 vs. NK-92 + VPA. (C) Immunohistochemical staining and quantification of MICA and MICB expression in the xenograft tumors; VPA significantly upregulated the expression of MICA and MICB in the pancreatic cancer xenograft tumors in vivo , this effect was attenuated by the PI3K inhibitor LY294002.

Article Snippet: Rabbit polyclonal antibodies against MICA and MICB were obtained from Santa Cruz, Santa Cruz, CA, USA.

Techniques: Lysis, In Vivo, Expressing, Immunohistochemical staining, Staining