Journal: Neurobiology of aging

Article Title: Prostate stem cell antigen interacts with nicotinic acetylcholine receptors and is affected in Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.01.001

Figure Lengend Snippet: Fig. 1. Prostate stem cell antigen (PSCA) is soluble and co-purifies with the a4 nAChR subunit in human cortex. (A) The antiserum directed against PSCA recognizes the human recombinant GST-tagged PSCA protein (total molecular weight 34.2 kDa) from 1 to 16 ng/well at the expected band size. Endogenous PSCA from mouse and human cortical tissue (8 mg total protein/well) is detected at approximately 24 kDa, and the amount in the cortical samples was compared with the known concentrations of re- combinant PSCA protein. (B) Images of Western blot showing PSCA protein levels in cortical tissue from human (protein concentration 4, 8, and 12 mg/well) and mouse (8, 15, and 30 mg/well) in the absence (peptide) and the presence of PSCA peptide (þpeptide). (C) Representative images of Western blots showing PSCA and Lypd6 protein levels in soluble and membrane fractions of human temporal cortical tissue. The membrane receptor proteins b2 nAChR and GluR2 are used as control. (D) Mag- netic beads covalently coupled with PSCA recombinant protein were incubated with cortical homogenates from human temporal cortex followed by detection of nAChR subunits by Western blot. Homogenates before (input) and after affinity purification (output) as well as the negative control (CTRL) were loaded.

Article Snippet: Validation of PSCA antibody (1:1,000, Novus Biologicals) was done with co-incubation with the PSCA peptide, which had been used as immunogen (1:100, # NB100-91938PEP, Novus Biologicals).

Techniques: Recombinant, Molecular Weight, Western Blot, Protein Concentration, Membrane, Control, Incubation, Negative Control

Journal: Genes, chromosomes & cancer

Article Title: Prostate stem cell antigen, a presumable organ-dependent tumor suppressor gene, is down-regulated in gallbladder carcinogenesis.

doi: 10.1002/gcc.20928

Figure Lengend Snippet: Figure 2. PSCA is expressed in normal gallbladder epithelium but down-regulated in GBCs. (A–E) Immunohistochemical double staining for PSCA (blue) and proliferating cell nuclear antigen (PCNA) (brown, for nuclear staining) proteins on gallbladder pathological specimens. The intensity was scored in five grades ‘‘111,’’ ‘‘11,’’ ‘‘1,’’ ‘‘1/2,’’ and ‘‘2/2.’’ A: normal gallbladder epithelium, B: papillary adenocarcinoma, C: well-differentiated adenocarcinoma, D: moder- ately differentiated adenocarcinoma, E: poorly differentiated adenocar- cinoma. (F) Immunohistochemical double staining of PSCA (blue) and PCNA (brown) in a poorly differentiated adenocarcinoma specimen shows PSCA expression in stromal cells (arrows). (G) Positive stain- ing of alpha-smooth muscle actin (a-SMA, blue) indicates the stromal cells are smooth muscle cells (arrowheads, double staining for PCNA). The specimen in G was from adjacent section of the speci- men in F obtained by serial sectioning. Bar: 100 lm. Arrows: PSCA signal in smooth muscle cells. (H) Quantitative RT-PCR analysis showed PSCA down-regulation in GBC tissues (T1–T4). The nontu- mor portion sample (N1) with the highest PSCA expression in the non-tumor counterparts of the four GBC specimens was used as reference.

Article Snippet: The sections were incubated at 4 C overnight with the mouse anti- PSCA antibody and rabbit anti-PCNA antibody (sc-7907, Santa Cruz, CA) simultaneously, and then with alkaline phosphatase conjugated antimouse IgG antibody and peroxidase conjugated anti-rabbit IgG antibody for 1 hr at room temperature.

Techniques: Immunohistochemical staining, Double Staining, Staining, Expressing, Quantitative RT-PCR

Journal: Genes, chromosomes & cancer

Article Title: Prostate stem cell antigen, a presumable organ-dependent tumor suppressor gene, is down-regulated in gallbladder carcinogenesis.

doi: 10.1002/gcc.20928

Figure Lengend Snippet: Figure 3. Immunohistochemical study revealed PSCA down-regu- lation in some chronic inflammatory conditions of gallbladder and in some rare types of GBC. Immunohistochemical double staining of PSCA (blue) and PCNA (brown) was performed. (A–D) chronic inflammatory conditions of gallbladder. A: xanthogranulomatous cho- lecystitis, B: chronic cholecystitis, C: adenomyomatosis, D: metaplasia. (E–G) rare types of GBC. E: mucinous adenocarcinoma, F: adeno- squamous cell carcinoma, G: undifferentiated carcinoma. Bar: 100 lm. Arrows: PSCA signal in smooth muscle cells.

Article Snippet: The sections were incubated at 4 C overnight with the mouse anti- PSCA antibody and rabbit anti-PCNA antibody (sc-7907, Santa Cruz, CA) simultaneously, and then with alkaline phosphatase conjugated antimouse IgG antibody and peroxidase conjugated anti-rabbit IgG antibody for 1 hr at room temperature.

Techniques: Immunohistochemical staining, Double Staining

Journal: Biology of reproduction

Article Title: Identification of distinct populations of prostasomes that differentially express prostate stem cell antigen, annexin A1, and GLIPR2 in humans.

doi: 10.1095/biolreprod.111.095760

Figure Lengend Snippet: FIG. 1. Fractionation by column chromatography. Membrane vesicles were collected from seminal plasma by ultracentrifugation and then separated by Sephacryl S-1000 column chromatography. A) CD9 and PSCA immunoblots of protein-containing fractions. Molecular weight markers are indicated in Mr 3 103. B and C) Representative whole mount transmission electron micrographs of membrane vesicles from pooled void volume fractions (28–34 from A, presented in B) and retained membrane vesicles (pooled fractions 44–54 from A, presented in C). Bars ¼ 500 nm. The data shown are representative of three independent experiments.

Article Snippet: Other blots were probed with mouse anti-human PSCA (clone 7F5; 1:1000; Santa Cruz Biotechnology Inc.), rabbit anti-human PSCA (1:250; Acris Antibodies Inc.), rabbit anti-human GLIPR2 [29] (1:5000), or mouse anti-human annexin A1 (clone 29; 1:250; BD Biosciences) [30].

Techniques: Fractionation, Column Chromatography, Membrane, Clinical Proteomics, Western Blot, Molecular Weight, Transmission Assay

Journal: Biology of reproduction

Article Title: Identification of distinct populations of prostasomes that differentially express prostate stem cell antigen, annexin A1, and GLIPR2 in humans.

doi: 10.1095/biolreprod.111.095760

Figure Lengend Snippet: FIG. 2. Fractionation on sucrose density gradients. Membrane vesicles from the pooled column fractions 44–54 in Figure 1A were loaded either on top (A) or at the bottom (B) of sucrose gradients and centrifuged for 62 h. C) Membrane vesicles were loaded at the bottom of a linear sucrose gradient and centrifuged for 16 h. CD9 and PSCA in gradient fractions were detected by immunoblotting as indicated. Molecular weight markers are indicated (Mr3103) on the right of the image, and the densities of the fractions (g/ml) are shown below the blots. The data shown are representative of six independent experiments.

Article Snippet: Other blots were probed with mouse anti-human PSCA (clone 7F5; 1:1000; Santa Cruz Biotechnology Inc.), rabbit anti-human PSCA (1:250; Acris Antibodies Inc.), rabbit anti-human GLIPR2 [29] (1:5000), or mouse anti-human annexin A1 (clone 29; 1:250; BD Biosciences) [30].

Techniques: Fractionation, Membrane, Western Blot, Molecular Weight

Journal: Biology of reproduction

Article Title: Identification of distinct populations of prostasomes that differentially express prostate stem cell antigen, annexin A1, and GLIPR2 in humans.

doi: 10.1095/biolreprod.111.095760

Figure Lengend Snippet: FIG. 3. Membrane vesicles from high- and low-density fractions are distinct in size but both contain CD9 and PSCA. Two vesicle populations were separated by sucrose gradient fractionation as indicated in Figure 2C. Vesicles from high- and low-density fractions were immobilized, immunogold- labeled for CD9 or PSCA, and analyzed by whole mount transmission electron microscopy. Representative pictures from two independent experiments are shown. Labeled vesicles are indicated by arrows. Bars¼ 500 nm. Vesicle mean sizes and percentages of vesicles labeled for CD9 or PSCA are indicated in Table 1.

Article Snippet: Other blots were probed with mouse anti-human PSCA (clone 7F5; 1:1000; Santa Cruz Biotechnology Inc.), rabbit anti-human PSCA (1:250; Acris Antibodies Inc.), rabbit anti-human GLIPR2 [29] (1:5000), or mouse anti-human annexin A1 (clone 29; 1:250; BD Biosciences) [30].

Techniques: Membrane, Fractionation, Labeling, Transmission Assay, Electron Microscopy

Journal: Biology of reproduction

Article Title: Identification of distinct populations of prostasomes that differentially express prostate stem cell antigen, annexin A1, and GLIPR2 in humans.

doi: 10.1095/biolreprod.111.095760

Figure Lengend Snippet: FIG. 4. PSCA is absent on prostasomes from some individual donors. Prostasomes were collected from the seminal fluid of individual donors (1–12) and immunoblotted for CD9 (A). The samples were probed on parallel blots for PSCA using mouse anti-human PSCA (B) or rabbit anti- human PSCA (C). Molecular weight markers are indicated on the right (Mr 3 103).

Article Snippet: Other blots were probed with mouse anti-human PSCA (clone 7F5; 1:1000; Santa Cruz Biotechnology Inc.), rabbit anti-human PSCA (1:250; Acris Antibodies Inc.), rabbit anti-human GLIPR2 [29] (1:5000), or mouse anti-human annexin A1 (clone 29; 1:250; BD Biosciences) [30].

Techniques: Molecular Weight

Journal: Neurobiology of aging

Article Title: Prostate stem cell antigen interacts with nicotinic acetylcholine receptors and is affected in Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.01.001

Figure Lengend Snippet: Fig. 1. Prostate stem cell antigen (PSCA) is soluble and co-purifies with the a4 nAChR subunit in human cortex. (A) The antiserum directed against PSCA recognizes the human recombinant GST-tagged PSCA protein (total molecular weight 34.2 kDa) from 1 to 16 ng/well at the expected band size. Endogenous PSCA from mouse and human cortical tissue (8 mg total protein/well) is detected at approximately 24 kDa, and the amount in the cortical samples was compared with the known concentrations of re- combinant PSCA protein. (B) Images of Western blot showing PSCA protein levels in cortical tissue from human (protein concentration 4, 8, and 12 mg/well) and mouse (8, 15, and 30 mg/well) in the absence (peptide) and the presence of PSCA peptide (þpeptide). (C) Representative images of Western blots showing PSCA and Lypd6 protein levels in soluble and membrane fractions of human temporal cortical tissue. The membrane receptor proteins b2 nAChR and GluR2 are used as control. (D) Mag- netic beads covalently coupled with PSCA recombinant protein were incubated with cortical homogenates from human temporal cortex followed by detection of nAChR subunits by Western blot. Homogenates before (input) and after affinity purification (output) as well as the negative control (CTRL) were loaded.

Article Snippet: Validation of PSCA antibody (1:1,000, Novus Biologicals) was done with co-incubation with the PSCA peptide, which had been used as immunogen (1:100, # NB100-91938PEP, Novus Biologicals).

Techniques: Recombinant, Molecular Weight, Western Blot, Protein Concentration, Membrane, Control, Incubation, Negative Control

Journal: Scientific Reports

Article Title: PSCA rs2294008 polymorphism contributes to the decreased risk for cervical cancer in a Chinese population

doi: 10.1038/srep23465

Figure Lengend Snippet: Association between PSCA SNP rs2294008 and risk of cervical cancer.

Article Snippet: Series of paraffin sections of specimens were incubated with polyclonal rabbit anti-PSCA antibody (BA1694, BOSTER, Wuhan, China) overnight at 4 °C, and 3,3′-diaminobenzidine (DAB; Zhongshan Biotech, Beijing, China) was used to produce a brown precipitate.

Techniques:

Journal: Scientific Reports

Article Title: PSCA rs2294008 polymorphism contributes to the decreased risk for cervical cancer in a Chinese population

doi: 10.1038/srep23465

Figure Lengend Snippet: Upper panel, representative images were obtained at 400× magnification. The frequency distribution of the CC, CT, and TT genotypes of rs2294008 was 25, 19, and 6, respectively. Bottom, the histogram of the PSCA expression in each genotype. ** P < 0.01.

Article Snippet: Series of paraffin sections of specimens were incubated with polyclonal rabbit anti-PSCA antibody (BA1694, BOSTER, Wuhan, China) overnight at 4 °C, and 3,3′-diaminobenzidine (DAB; Zhongshan Biotech, Beijing, China) was used to produce a brown precipitate.

Techniques: Expressing

Journal: Biomedicines

Article Title: Differential Protein-Coding Gene Expression Profile in Patients with Prostate Cancer

doi: 10.3390/biomedicines12112509

Figure Lengend Snippet: Genes classified as drug targets in the DrugBank database. Protein-coding genes as targets for drugs related to the treatment of prostate cancer in all states (investigation, experimental, approved). In the case of target genes for many drugs, only the first 10 are reported.

Article Snippet: DB05933 , MK-4721 , A fully human monoclonal antibody directed to Agensys’ proprietary target Prostate Stem Cell Antigen (PSCA), an antigen expressed at significant levels on tumor cells from the majority of patients with all stages of prostate, pancreatic, and bladder cancers [investigation]. , PSCA [O43653].

Techniques: Diagnostic Assay, Positron Emission Tomography, Membrane, Imaging, Biomarker Discovery, Expressing, Derivative Assay

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Prostate stem cell antigen (PSCA) expression in primary GC tissues and cell lines. a. Immunohistochemical staining for PSCA in normal gastric tissue and eight primary GC samples; scale bar = 100 μm. b. Detection of PSCA expression in three human GC cell lines, BGC-823, KATO III, and MKN-28 cells, by flow cytometry

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Expressing, Immunohistochemical staining, Staining, Flow Cytometry

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Generation of anti-prostate stem cell antigen (PSCA) CAR-T cells. a . The discrete CAR units of anti-PSCA CAR-T cells and GFP-T cells. b . Representative flow cytometric analyses of transfected T cells detected by flow cytometry. c . CCR7, CD62L, CD45RA, and CD45RO expression was detected on T cells after their generation

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Transfection, Flow Cytometry, Expressing

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Anti-PSCA CAR-T cells exhibited dramatic antitumor efficacy ex vivo. a . The lytic capacity towards different target cells, including BGC-823, KATO III and MKN-28 cells, was analyzed at the indicated effector-to-target ratios in a 24 h lysis assay. b . The concentrations of IL-2, IFN-γ, GM-CSF, and TNFα released by anti-PSCA CAR-T cells and GFP-T cells after coculture with BGC-823 cells overnight at an E:T ratio of 1:1 are shown. Error bars denote the s.e.m., and the results were calculated by an unpaired t test. * indicates p < 0.05; ** indicates p < 0.01; and *** indicates p < 0.001. c . Canonical T cell markers were detected by flow cytometry at a recommended E:T ratio of 1:1 after coculturing anti-PSCA CAR-T cells and GFP- T cells with target cell lines. d . Statistical analysis of three independent FACS results. Error bars denote the s.e.m.

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Ex Vivo, Lysis, Flow Cytometry

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Anti-PSCA CAR-T cells efficiently reduced tumor progression in BGC-823 models. a . Schematic representation depicting the time course of the experiment. b . Tumor volume was calculated according to the following formula: length × width 2 /2. c . Tumor weight of BGC-823 subcutaneously injected mice. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. d . Percentage of T cells in the tumor of the BGC-823 models. e . Percentage of T cells in the PB of the BGC-823 models and representative FACS plots of the i.v group and the p.t group, respectively. f . Percentage of T cells in the spleen of the BGC-823 models and representative FACS plots of the i.v group and the p.t group, respectively. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Injection

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Anti-PSCA CAR-T cells suppressed tumor progression in MKN-28 models. a . Schematic representation depicting the time course of the experiment. b . Tumor volume was calculated according to the following formula: length × width 2 /2. c . Tumor weight of MKN-28 subcutaneously injected mice. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. d . Percentage of T cells in the tumor of the MKN-28 models. e . Percentage of T cells in the PB of the MKN-28 models and representative FACS plots of the i.v group and the p.t group, respectively. f . Percentage of T cells in the spleen of the MKN-28 models and representative FACS plots of the i.v group and the p.t group, respectively. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Injection