Journal: Oncology Reports

Article Title: MicroRNA-195-5p is associated with cell proliferation, migration and invasion in prostate cancer and targets MIB1

doi: 10.3892/or.2021.8210

Figure Lengend Snippet: Primer sequences for reverse transcription-quantitative PCR.

Article Snippet: The recombinant plasmid vectors expressing the double-stranded shRNA targeting MIB1 (sh-MIB1) or the shRNA control (sh-NC) were purchased from Shanghai GenePharma Co., Ltd.

Techniques: Sequencing

Journal: Oncology Reports

Article Title: MicroRNA-195-5p is associated with cell proliferation, migration and invasion in prostate cancer and targets MIB1

doi: 10.3892/or.2021.8210

Figure Lengend Snippet: MIB1 is highly expressed in PCa tissues and cell lines. The mRNA expression level of MIB1 was measured in (A) PCa tissues and adjacent normal tissues (n=40), and in (B) PCa cell lines (PC-3, VCaP, 22Rv1, DU145 and LNCaP) and a normal human prostate epithelial cell line (RWPE1) using reverse transcription-quantitative PCR analysis. ***P<0.001. (C) Western blot analysis of MIB1 protein expression level in PCa cell lines (PC-3, VCaP, 22Rv1, DU145 and LNCaP) and a normal human prostate epithelial cell line (RWPE1). MIB1, mind bomb 1; PCa, prostate cancer.

Article Snippet: The recombinant plasmid vectors expressing the double-stranded shRNA targeting MIB1 (sh-MIB1) or the shRNA control (sh-NC) were purchased from Shanghai GenePharma Co., Ltd.

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

Journal: Oncology Reports

Article Title: MicroRNA-195-5p is associated with cell proliferation, migration and invasion in prostate cancer and targets MIB1

doi: 10.3892/or.2021.8210

Figure Lengend Snippet: MIB1 promotes PCa cell proliferation, migration and invasion. To elucidate the function of MIB1 in PCa, the VCaP and DU145 cells were transfected with OE-MIB1, sh-MIB1, OE-NC or sh-NC. (A) mRNA and protein expression level of MIB1 in the transfected VCaP and DU145 cells was measured using reverse transcription-quantitative PCR and western blot analysis, respectively. (B) Cell Counting Kit-8 assay was used to assess the proliferation of the transfected VCaP and DU145 cell lines. Transwell assays were used to evaluate the (C) migration and (D) invasion of the transfected VCaP and DU145 cell lines. *P<0.05, **P<0.01 and ***P<0.001. MIB1, mind bomb 1; PCa, prostate cancer; OE, overexpression; sh, short hairpin RNA; NC, negative control; OD, optical density.

Article Snippet: The recombinant plasmid vectors expressing the double-stranded shRNA targeting MIB1 (sh-MIB1) or the shRNA control (sh-NC) were purchased from Shanghai GenePharma Co., Ltd.

Techniques: Migration, Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Counting, Over Expression, shRNA, Negative Control

Journal: Oncology Reports

Article Title: MicroRNA-195-5p is associated with cell proliferation, migration and invasion in prostate cancer and targets MIB1

doi: 10.3892/or.2021.8210

Figure Lengend Snippet: miR-195-5p directly interacts with MIB1. (A) Putative binding sites between miR-195-5p and MIB1, and the mutant sequences of MIB1. Luciferase activity was detected in (B) VCaP and (C) DU145 cells co-transfected with MIB1-WT or MIB1-MUT, and miR-195-5p mimics or miR-NC. **P<0.01. miR, microRNA; MIB1, mind bomb 1; WT, wild-type; MUT, mutant; NC, negative control; UTR, untranslated region.

Article Snippet: The recombinant plasmid vectors expressing the double-stranded shRNA targeting MIB1 (sh-MIB1) or the shRNA control (sh-NC) were purchased from Shanghai GenePharma Co., Ltd.

Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Transfection, Negative Control

Journal: Oncology Reports

Article Title: MicroRNA-195-5p is associated with cell proliferation, migration and invasion in prostate cancer and targets MIB1

doi: 10.3892/or.2021.8210

Figure Lengend Snippet: miR-195-5p expression level is elevated in PCa tissues and cell lines, and MIB1 is directly regulated by miR-195-5p. The mRNA expression level of miR-195-5p was examined using RT-qPCR in (A) PCa tissues and adjacent normal tissues (n=40), as well as in (B) PCa cell lines (PC-3, VCaP, 22Rv1, DU145 and LNCaP) and a normal human prostate epithelial cell line (RWPE1). (C) Pearson's analysis was used to assess the correlation between MIB1 and miR-195-5p expression in PCa. (D) MIB1 expression was measured using RT-qPCR and western blot anlaysis in VCaP and DU145 cells transfected with miR-195-5p mimics, anti-miR-195-5p, miR-NC or anti-miR-NC. *P<0.05, **P<0.01 and ***P<0.001. miR, microRNA; PCa, prostate cancer; MIB1, mind bomb 1; RT-qPCR, reverse transcription-quantitative PCR; NC, negative control.

Article Snippet: The recombinant plasmid vectors expressing the double-stranded shRNA targeting MIB1 (sh-MIB1) or the shRNA control (sh-NC) were purchased from Shanghai GenePharma Co., Ltd.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Real-time Polymerase Chain Reaction, Negative Control

Journal: Oncology Reports

Article Title: MicroRNA-195-5p is associated with cell proliferation, migration and invasion in prostate cancer and targets MIB1

doi: 10.3892/or.2021.8210

Figure Lengend Snippet: miR-195-5p promotes the proliferation and invasion of PCa cells via MIB1. To elucidate the function of the miR-195-5p/MIB1 axis in PCa, the VCaP and DU145 cell lines were transfected with miR-195-5p mimics, miR-NC, anti-miR-195-5p, anti-miR-NC, miR-195-5p mimics + OE-NC, miR-195-5p mimics + OE-MIB1, anti-miR-195-5p + sh-MIB1 or anti-miR-195-5p + sh-NC. The mRNA and protein expression levels of MIB1 was determined in the VCaP and DU145 cell lines using reverse transcription-quantitative PCR and western blot analysis, respectively following transfection with (A) miR-195-5p mimics and OE-MIB1, and (B) anti-miR-195-5p and sh-MIB1. Cell Counting Kit-8 assay was used to assess the proliferation of VCaP and DU145 cells transfected with (C) miR-195-5p mimics, miR-NC, miR-195-5p mimics + OE-NC, miR-195-5p mimic + OE-MIB1, and (D) anti-miR-195-5p, anti-miR-NC, anti-miR-195-5p + sh-MIB1 or anti-miR-195-5p + sh-NC. Transwell assay was used to detect the invasion of VCaP and DU145 cells transfected with (E) miR-195-5p mimics, miR-NC, miR-195-5p mimics + OE-NC, miR-195-5p mimics + OE-MIB1 and (F) anti-miR-195-5p, anti-miR-NC, anti-miR-195-5p + sh-MIB1 or anti-miR-195-5p + sh-NC. *P<0.05, **P<0.01 and ***P<0.001. miR, microRNA; PCa, prostate cancer; MIB1, mind bomb 1; NC, negative control; OE, overexpression; sh, short hairpin RNA; OD, optical density.

Article Snippet: The recombinant plasmid vectors expressing the double-stranded shRNA targeting MIB1 (sh-MIB1) or the shRNA control (sh-NC) were purchased from Shanghai GenePharma Co., Ltd.

Techniques: Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Counting, Transwell Assay, Negative Control, Over Expression, shRNA

Journal: British Journal of Cancer

Article Title: Anti-glypican-1 antibody–drug conjugate is a potential therapy against pancreatic cancer

doi: 10.1038/s41416-020-0781-2

Figure Lengend Snippet: a Representative images of IHC staining for GPC in pancreatic cancer specimens. GPC-1 expression in cell membranes of human pancreatic cancer. Left panel: the high GPC-1 expression group (HG); right panel: the low GPC-1 expression group (LG). Scale bar: 50 μm. b The distribution of GPC-1 scores in LG and HG. c Kaplan–Meier analyses of OS. The black line represents LG and the grey line represents HG. d Kaplan–Meier analyses of RFS.

Article Snippet: To generate a stable GPC-1-knockdown cell line, BxPC-3 cells were transfected with a commercial plasmid vector expressing short-hairpin RNA (shRNA) targeting GPC-1 mRNA or a negative nonspecific shRNA control (SuperArray Bioscience Corp., Frederick, MD, USA) using Lipofectamine TM 2000 (Life Technologies Corp., Carlsbad, CA, USA).

Techniques: Immunohistochemistry, Expressing

Journal: British Journal of Cancer

Article Title: Anti-glypican-1 antibody–drug conjugate is a potential therapy against pancreatic cancer

doi: 10.1038/s41416-020-0781-2

Figure Lengend Snippet: a Flow cytometry of GPC-1 expression in BxPC-3, T3M-4 and SUIT-2 using anti-GPC-1 monoclonal antibody. b BxPC-3, T3M-4 and SUIT-2 cells were treated with anti-GPC-1 monoclonal antibody (clone 01a033) or control IgG antibody for 144 h. Neither antibody inhibited the growth of any cell line. c Cells were treated with GPC-1-ADC or mouse IgG2a-ADC (a control ADC) for 144 h. Relative to control ADC, GPC-1-ADC significantly inhibited the growth of GPC-1-positive BxPC-3 and T3M-4 cell lines. Neither treatment inhibited the growth of the GPC-1-negative SUIT-2 cell line. d Time course of the internalisation activity of GPC-1-ADC in BxPC-3 and T3M-4 cells. e GPC-1-ADC internalises and locates in the lysosomes of BxPC-3 cells. Plasma membrane and intracellular GPC-1 were visualised by confocal fluorescence microscopy. Green indicates GPC-1-ADC, red indicates the lysosomal marker LAMP-1 and blue indicates DAPI (4′,6-diamidino-2-phenylindole)-stained DNA. Scale bar: 10 μm. GPC-1-ADC was found in the lysosomes. GPC-1-ADC and the lysosomal marker LAMP-1 overlapped (arrow).

Article Snippet: To generate a stable GPC-1-knockdown cell line, BxPC-3 cells were transfected with a commercial plasmid vector expressing short-hairpin RNA (shRNA) targeting GPC-1 mRNA or a negative nonspecific shRNA control (SuperArray Bioscience Corp., Frederick, MD, USA) using Lipofectamine TM 2000 (Life Technologies Corp., Carlsbad, CA, USA).

Techniques: Flow Cytometry, Expressing, Activity Assay, Fluorescence, Microscopy, Marker, Staining

Journal: British Journal of Cancer

Article Title: Anti-glypican-1 antibody–drug conjugate is a potential therapy against pancreatic cancer

doi: 10.1038/s41416-020-0781-2

Figure Lengend Snippet: a Flow cytometry of GPC-1 expression in a GPC-1-knockdown cell line (BxPC-3 KD-2-23) and a control cell line (BxPC-3 NC-11) using anti-GPC-1 monoclonal antibody. b Quantitative reverse transcription-PCR analysis of GPC-1 mRNA levels relative to β-actin in a GPC-1-knockdown cell line (BxPC-3 KD-2-23) and control cell lines (BxPC-3 and BxPC-3 NC-11). c BxPC-3, BxPC-3 NC-11 and BxPC-3 KD-2-23 were treated with GPC-1-ADC for 144 h. GPC-1-ADC had a lower growth inhibition effect of the GPC-1-knockdown cell line (BxPC-3 KD-2-23) than the parent BxPC-3 and the negative control cell line (BxPC-3 NC-11).

Article Snippet: To generate a stable GPC-1-knockdown cell line, BxPC-3 cells were transfected with a commercial plasmid vector expressing short-hairpin RNA (shRNA) targeting GPC-1 mRNA or a negative nonspecific shRNA control (SuperArray Bioscience Corp., Frederick, MD, USA) using Lipofectamine TM 2000 (Life Technologies Corp., Carlsbad, CA, USA).

Techniques: Flow Cytometry, Expressing, Inhibition, Negative Control

Journal: British Journal of Cancer

Article Title: Anti-glypican-1 antibody–drug conjugate is a potential therapy against pancreatic cancer

doi: 10.1038/s41416-020-0781-2

Figure Lengend Snippet: a Antitumour efficacy of GPC-1-ADC in BxPC-3 xenograft models ( n = 8 or 9). Representative images of IHC staining for GPC-1 in xenografted tumour tissues from untreated mice. Tumour-bearing mice were intravenously administered PBS, control ADC (10 mg kg –1 ) or GPC-1-ADC (1 mg kg –1 , 3 mg kg –1 or 10 mg kg –1 ) on days 0, 4, 8 and 12. Each point on the graph represents the average tumour volume. b Thirty-six days after the first treatment, the tumour weights were calculated. c . Changes in body weight are represented. d GPC-1-ADC causes mitotic arrest in vivo. Animals with BxPC-3 tumour xenografts were administered a single dose of PBS, control ADC (10 mg kg –1 ) or GPC-1-ADC (1 mg kg –1 , 3 mg kg –1 or 10 mg kg –1 ). After 24 h, the tumours were harvested and stained with anti-phospho-histone H3 (Ser10) antibody to detect mitotic cells. Scale bar: 200 μm. e . Phospho-histone H3 (Ser10) staining was recorded as the ratio of positively stained cells to all tumour cells in five fields (×200 magnification).

Article Snippet: To generate a stable GPC-1-knockdown cell line, BxPC-3 cells were transfected with a commercial plasmid vector expressing short-hairpin RNA (shRNA) targeting GPC-1 mRNA or a negative nonspecific shRNA control (SuperArray Bioscience Corp., Frederick, MD, USA) using Lipofectamine TM 2000 (Life Technologies Corp., Carlsbad, CA, USA).

Techniques: Immunohistochemistry, In Vivo, Staining

Journal: British Journal of Cancer

Article Title: Anti-glypican-1 antibody–drug conjugate is a potential therapy against pancreatic cancer

doi: 10.1038/s41416-020-0781-2

Figure Lengend Snippet: a Antitumour efficacy of GPC-1-ADC in patient-derived xenograft models ( n = 6). Representative images of IHC staining for GPC-1 in xenografted tumour tissues from untreated mice. Tumour-bearing mice were intravenously administered PBS, control ADC (10 mg kg –1 ) or GPC-1-ADC (1 mg kg –1 , 3 mg kg –1 or 10 mg kg –1 ) on days 0, 4, 8 and 12. Each point on the graph represents the average tumour volume. b Twenty-eight days after the first treatment, the tumour weights were calculated. c Changes in body weight are represented. d GPC-1-ADC causes mitotic arrest in vivo. Animals with patient-derived xenografts were administered a single dose of PBS, control ADC (10 mg kg –1 ) or GPC-1-ADC (10 mg kg –1 ). After 24 h, tumours were harvested and stained with anti-phospho-histone H3 (Ser10) antibody to detect mitotic cells. Scale bar: 200 μm. e Phospho-histone H3 (Ser10) staining was recorded as the ratio of positively stained cells to all tumour cells in five fields (×200 magnification).

Article Snippet: To generate a stable GPC-1-knockdown cell line, BxPC-3 cells were transfected with a commercial plasmid vector expressing short-hairpin RNA (shRNA) targeting GPC-1 mRNA or a negative nonspecific shRNA control (SuperArray Bioscience Corp., Frederick, MD, USA) using Lipofectamine TM 2000 (Life Technologies Corp., Carlsbad, CA, USA).

Techniques: Derivative Assay, Immunohistochemistry, In Vivo, Staining