Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: TNF-α modulates the WISP1 expression in stroma cells of the human bladder. (A) The RT-qPCR determined the expression of WISP1 in the human bladder cells. (B) Two WISP1 isoforms (WISP1v1 and WISP1v2) were found in the human bladder fibroblast (HBdSF) and bladder smooth muscle (HBdSMC) cells by RT-PCR with the pair of primers as shown. The expressions of WISP1, IL-6, CXCL5, CXCL12, and GDF15 of the HBdSF (C) and HBdSMC (D) cells after 10 ng/ml TNF-α treatment assessed by RT-qPCR. WISP1 (E) and IL-6 (F) secretions after TNF-α (0 – 20 ng/mL) treatments in HBdSF cells. Secretion of WISP1 (G) and IL-6 (F) in HBdSMC_shCOL and HBdSMC_shWISP1 cells treated with/without 10 ng/ml of TNF-α. Data are presented as the mean percentage (± SE; n = 4) in relation to the vehicle-treated HBdSMC_shCOL cells. (I) WISP1 secretion after 20 μM CAPE and/or TNF-α in HBdSF cells determined by ELISA assays. Data are presented as the mean percentage (±SE; n = 4) in relation to the vehicle-treated HBdSF cells. (J) The luciferase activity of the WISP1 reporter vector when HBdSMC cells were treated with TNF-α (10 ng/mL) and/or 20 μM CAPE for 24 h. * p < 0.05; ** p < 0.01; ND: no detectable.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Plasmid Preparation

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of the conditioned media from the ectopic WISP1 overexpressed-293T cells on the cell proliferation in the bladder stroma cells. Expressions of WISP1v1 and WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the 293T cells were assessed by the RT-qPCR (A) and ELISA (B) assays. The ability of the proliferation of HBdSMC (C, D) and HBdSF (E, F) cells after being treated with the supernatant from 293T-DNA, 293T-WISP1v1, and 293T-WISP1v2 cells, respectively, was measured by the EdU staining or EdU flow cytometry assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. * p < 0.05; ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: WISP1 modulates expressions of α-SMA, IL-6, GDF15, and CXCL5 in bladder stroma cells. The mRNA levels of two WISP1 isoforms (WISP1v1 and WISP1v2) after WISP1 knock-downed in the HBdSF (A) and HBdSMC (B) cells were determined by RT-PCR. The protein levels with quantification analysis of WISP1, GDF15, and α-SMA of HBdSMC (C) and HBdSF (D) cells after mock-knockdown or WISP1-knockdown were assessed by immunoblot assays. The mRNA levels of the WISP1, α-SMA, IL-6, GDF15, and CXCL5 after knockdown of WISP1 in the HBdSMC (E) and HBdSF (F) cells, as indicated, were determined by RT-qPCR. Data are presented as target genes/β-actin of mock-knockdown cells relative to WISP1-knockdown cells. ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Reverse Transcription Polymerase Chain Reaction, Knockdown, Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1-knockdown on the cell proliferation, migration, and contraction in bladder stroma and cancer cells. (A) The ability of cell proliferation of mock-knockdown HBdSMC (HBdSMC_shCOL) and WISPI-knockdown (HBdSMC_shWISP1) cells was assessed by EdU proliferation assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. (B) The ability of cell contraction of the mock-knockdown HBdSF (HBdSF_shCOL) and the WISP1-knockdown HBdSF (HBdSF_shWISP1) cells was measured by the collage contraction assays. Data are presented as gel area of the times as indicated in relation to the 0 h. (C) The ability of the migration of T24 cells after being treated with the supernatant from HBdSMC_shCOL and HBdSMC_shWISP1 cells, respectively, was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Knockdown, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1 on the cell proliferation and invasion in bladder cancer cells. (A) Cell proliferation of HT1376 and T24 cells after treatments of recombinant human WISP1 was assessed by CyQUANT cell proliferation assay. Data are presented as the mean percentage of the rhWISP1-treated cells relative to vehicle-treated cells. Expressions of total WISP1, WISP1v1, or WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the HT1376 cells were assessed by RT-qPCR (B), RT-PCR (C), and ELISA (D) assays. The abilities of the proliferation of HT-DNA, HT-WISP1v1 (E), and HT-WISP1v2 (F) cells were measured by Ki67 flow cytometry assays. Data are presented as the mean percentage of the Ki67-positive cells relative to HT-DNA cells. The invasion ability of HT1376 cells after ectopic overexpression of WISP1v1 (G) and WISP1v2 (H) was determined by in vitro Matrigel invasion assays (± SE; n = 4). Data are presented as the mean percentage of the invasion cells relative to HT-DNA cells. * p < 0.05; ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Recombinant, CyQUANT Assay, Proliferation Assay, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Over Expression, In Vitro

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Determination of the molecular mechanism of WISP1v1 on the cell invasion in the bladder cancer cells. (A) The protein levels of NDRG1, KAI1, and Maspin of HT-DNA and HT-WISP1v1 cells were determined by immunoblot assays. (B) The luciferase activity of the NDRG1 reporter vector was shown after transiently overexpressing various amounts of WISP1v1 expression vectors in HT1376 and T24 cells, as indicated. (C) The luciferase activity of KAI1 and Maspin reporter vectors was shown as indicated after transiently overexpressing various dosages of WISP1v1 expression vectors in HT1376 cells. (D) The luciferase activity of NDRG1, KAI1, and Maspin reporter vectors, respectively, after transiently overexpressed pcDNA, WISP1v1, and WISP1v2 expression vectors as indicated in HT1376 cells. (E) The mRNA levels of the E-cadherin, N-cadherin, Slug, Snail, and Vimentin were determined by RT-qPCR. Data are presented as target genes/β-actin of HT-WISP1v1 cells relative to HT-DNA cells. The mRNA levels of the WISP1 (F), NDRG1, KAI1, Maspin, and IL-6 (G) after transient overexpression of WISP1v1 in T24 cells were determined by RT-qPCR. Data are presented as target genes/β-actin of T24-WISP1v1 cells relative to T24-DNA cells. (H) The invasion ability of T24 cells after the ectopic overexpression of WISP1v1 was determined by in vitro Matrigel invasion assays (± SE; n = 4). The ability of the migration (I) and quantification analysis (I) of T24 cells after being treated with the supernatant from 293T-DNA and 293T-WISP1v1 cells was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. * p < 0.05, ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Western Blot, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Quantitative RT-PCR, Over Expression, In Vitro, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: TNF-α modulates the WISP1 expression in stroma cells of the human bladder. (A) The RT-qPCR determined the expression of WISP1 in the human bladder cells. (B) Two WISP1 isoforms (WISP1v1 and WISP1v2) were found in the human bladder fibroblast (HBdSF) and bladder smooth muscle (HBdSMC) cells by RT-PCR with the pair of primers as shown. The expressions of WISP1, IL-6, CXCL5, CXCL12, and GDF15 of the HBdSF (C) and HBdSMC (D) cells after 10 ng/ml TNF-α treatment assessed by RT-qPCR. WISP1 (E) and IL-6 (F) secretions after TNF-α (0 – 20 ng/mL) treatments in HBdSF cells. Secretion of WISP1 (G) and IL-6 (F) in HBdSMC_shCOL and HBdSMC_shWISP1 cells treated with/without 10 ng/ml of TNF-α. Data are presented as the mean percentage (± SE; n = 4) in relation to the vehicle-treated HBdSMC_shCOL cells. (I) WISP1 secretion after 20 μM CAPE and/or TNF-α in HBdSF cells determined by ELISA assays. Data are presented as the mean percentage (±SE; n = 4) in relation to the vehicle-treated HBdSF cells. (J) The luciferase activity of the WISP1 reporter vector when HBdSMC cells were treated with TNF-α (10 ng/mL) and/or 20 μM CAPE for 24 h. * p < 0.05; ** p < 0.01; ND: no detectable.

Article Snippet: Cells were transduced with WISP1 shRNA lentiviral transduction particles (sc-39,335-V; Santa Cruz Biotechnology, CA, USA) or control shRNA transduction particles (sc-10,808-v, Santa Cruz Biotechnology), and selected with 50 ng/mL puromycin dihydrochloride (Cyrusbioscience, Taipei, Taiwan) as described previously [ ].

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Plasmid Preparation

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of the conditioned media from the ectopic WISP1 overexpressed-293T cells on the cell proliferation in the bladder stroma cells. Expressions of WISP1v1 and WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the 293T cells were assessed by the RT-qPCR (A) and ELISA (B) assays. The ability of the proliferation of HBdSMC (C, D) and HBdSF (E, F) cells after being treated with the supernatant from 293T-DNA, 293T-WISP1v1, and 293T-WISP1v2 cells, respectively, was measured by the EdU staining or EdU flow cytometry assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. * p < 0.05; ** p < 0.01.

Article Snippet: Cells were transduced with WISP1 shRNA lentiviral transduction particles (sc-39,335-V; Santa Cruz Biotechnology, CA, USA) or control shRNA transduction particles (sc-10,808-v, Santa Cruz Biotechnology), and selected with 50 ng/mL puromycin dihydrochloride (Cyrusbioscience, Taipei, Taiwan) as described previously [ ].

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: WISP1 modulates expressions of α-SMA, IL-6, GDF15, and CXCL5 in bladder stroma cells. The mRNA levels of two WISP1 isoforms (WISP1v1 and WISP1v2) after WISP1 knock-downed in the HBdSF (A) and HBdSMC (B) cells were determined by RT-PCR. The protein levels with quantification analysis of WISP1, GDF15, and α-SMA of HBdSMC (C) and HBdSF (D) cells after mock-knockdown or WISP1-knockdown were assessed by immunoblot assays. The mRNA levels of the WISP1, α-SMA, IL-6, GDF15, and CXCL5 after knockdown of WISP1 in the HBdSMC (E) and HBdSF (F) cells, as indicated, were determined by RT-qPCR. Data are presented as target genes/β-actin of mock-knockdown cells relative to WISP1-knockdown cells. ** p < 0.01.

Article Snippet: Cells were transduced with WISP1 shRNA lentiviral transduction particles (sc-39,335-V; Santa Cruz Biotechnology, CA, USA) or control shRNA transduction particles (sc-10,808-v, Santa Cruz Biotechnology), and selected with 50 ng/mL puromycin dihydrochloride (Cyrusbioscience, Taipei, Taiwan) as described previously [ ].

Techniques: Reverse Transcription Polymerase Chain Reaction, Knockdown, Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1-knockdown on the cell proliferation, migration, and contraction in bladder stroma and cancer cells. (A) The ability of cell proliferation of mock-knockdown HBdSMC (HBdSMC_shCOL) and WISPI-knockdown (HBdSMC_shWISP1) cells was assessed by EdU proliferation assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. (B) The ability of cell contraction of the mock-knockdown HBdSF (HBdSF_shCOL) and the WISP1-knockdown HBdSF (HBdSF_shWISP1) cells was measured by the collage contraction assays. Data are presented as gel area of the times as indicated in relation to the 0 h. (C) The ability of the migration of T24 cells after being treated with the supernatant from HBdSMC_shCOL and HBdSMC_shWISP1 cells, respectively, was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. ** p < 0.01.

Article Snippet: Cells were transduced with WISP1 shRNA lentiviral transduction particles (sc-39,335-V; Santa Cruz Biotechnology, CA, USA) or control shRNA transduction particles (sc-10,808-v, Santa Cruz Biotechnology), and selected with 50 ng/mL puromycin dihydrochloride (Cyrusbioscience, Taipei, Taiwan) as described previously [ ].

Techniques: Knockdown, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1 on the cell proliferation and invasion in bladder cancer cells. (A) Cell proliferation of HT1376 and T24 cells after treatments of recombinant human WISP1 was assessed by CyQUANT cell proliferation assay. Data are presented as the mean percentage of the rhWISP1-treated cells relative to vehicle-treated cells. Expressions of total WISP1, WISP1v1, or WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the HT1376 cells were assessed by RT-qPCR (B), RT-PCR (C), and ELISA (D) assays. The abilities of the proliferation of HT-DNA, HT-WISP1v1 (E), and HT-WISP1v2 (F) cells were measured by Ki67 flow cytometry assays. Data are presented as the mean percentage of the Ki67-positive cells relative to HT-DNA cells. The invasion ability of HT1376 cells after ectopic overexpression of WISP1v1 (G) and WISP1v2 (H) was determined by in vitro Matrigel invasion assays (± SE; n = 4). Data are presented as the mean percentage of the invasion cells relative to HT-DNA cells. * p < 0.05; ** p < 0.01.

Article Snippet: Cells were transduced with WISP1 shRNA lentiviral transduction particles (sc-39,335-V; Santa Cruz Biotechnology, CA, USA) or control shRNA transduction particles (sc-10,808-v, Santa Cruz Biotechnology), and selected with 50 ng/mL puromycin dihydrochloride (Cyrusbioscience, Taipei, Taiwan) as described previously [ ].

Techniques: Recombinant, CyQUANT Assay, Proliferation Assay, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Over Expression, In Vitro

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Determination of the molecular mechanism of WISP1v1 on the cell invasion in the bladder cancer cells. (A) The protein levels of NDRG1, KAI1, and Maspin of HT-DNA and HT-WISP1v1 cells were determined by immunoblot assays. (B) The luciferase activity of the NDRG1 reporter vector was shown after transiently overexpressing various amounts of WISP1v1 expression vectors in HT1376 and T24 cells, as indicated. (C) The luciferase activity of KAI1 and Maspin reporter vectors was shown as indicated after transiently overexpressing various dosages of WISP1v1 expression vectors in HT1376 cells. (D) The luciferase activity of NDRG1, KAI1, and Maspin reporter vectors, respectively, after transiently overexpressed pcDNA, WISP1v1, and WISP1v2 expression vectors as indicated in HT1376 cells. (E) The mRNA levels of the E-cadherin, N-cadherin, Slug, Snail, and Vimentin were determined by RT-qPCR. Data are presented as target genes/β-actin of HT-WISP1v1 cells relative to HT-DNA cells. The mRNA levels of the WISP1 (F), NDRG1, KAI1, Maspin, and IL-6 (G) after transient overexpression of WISP1v1 in T24 cells were determined by RT-qPCR. Data are presented as target genes/β-actin of T24-WISP1v1 cells relative to T24-DNA cells. (H) The invasion ability of T24 cells after the ectopic overexpression of WISP1v1 was determined by in vitro Matrigel invasion assays (± SE; n = 4). The ability of the migration (I) and quantification analysis (I) of T24 cells after being treated with the supernatant from 293T-DNA and 293T-WISP1v1 cells was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were transduced with WISP1 shRNA lentiviral transduction particles (sc-39,335-V; Santa Cruz Biotechnology, CA, USA) or control shRNA transduction particles (sc-10,808-v, Santa Cruz Biotechnology), and selected with 50 ng/mL puromycin dihydrochloride (Cyrusbioscience, Taipei, Taiwan) as described previously [ ].

Techniques: Western Blot, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Quantitative RT-PCR, Over Expression, In Vitro, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: TNF-α modulates the WISP1 expression in stroma cells of the human bladder. (A) The RT-qPCR determined the expression of WISP1 in the human bladder cells. (B) Two WISP1 isoforms (WISP1v1 and WISP1v2) were found in the human bladder fibroblast (HBdSF) and bladder smooth muscle (HBdSMC) cells by RT-PCR with the pair of primers as shown. The expressions of WISP1, IL-6, CXCL5, CXCL12, and GDF15 of the HBdSF (C) and HBdSMC (D) cells after 10 ng/ml TNF-α treatment assessed by RT-qPCR. WISP1 (E) and IL-6 (F) secretions after TNF-α (0 – 20 ng/mL) treatments in HBdSF cells. Secretion of WISP1 (G) and IL-6 (F) in HBdSMC_shCOL and HBdSMC_shWISP1 cells treated with/without 10 ng/ml of TNF-α. Data are presented as the mean percentage (± SE; n = 4) in relation to the vehicle-treated HBdSMC_shCOL cells. (I) WISP1 secretion after 20 μM CAPE and/or TNF-α in HBdSF cells determined by ELISA assays. Data are presented as the mean percentage (±SE; n = 4) in relation to the vehicle-treated HBdSF cells. (J) The luciferase activity of the WISP1 reporter vector when HBdSMC cells were treated with TNF-α (10 ng/mL) and/or 20 μM CAPE for 24 h. * p < 0.05; ** p < 0.01; ND: no detectable.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Plasmid Preparation

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of the conditioned media from the ectopic WISP1 overexpressed-293T cells on the cell proliferation in the bladder stroma cells. Expressions of WISP1v1 and WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the 293T cells were assessed by the RT-qPCR (A) and ELISA (B) assays. The ability of the proliferation of HBdSMC (C, D) and HBdSF (E, F) cells after being treated with the supernatant from 293T-DNA, 293T-WISP1v1, and 293T-WISP1v2 cells, respectively, was measured by the EdU staining or EdU flow cytometry assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. * p < 0.05; ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: WISP1 modulates expressions of α-SMA, IL-6, GDF15, and CXCL5 in bladder stroma cells. The mRNA levels of two WISP1 isoforms (WISP1v1 and WISP1v2) after WISP1 knock-downed in the HBdSF (A) and HBdSMC (B) cells were determined by RT-PCR. The protein levels with quantification analysis of WISP1, GDF15, and α-SMA of HBdSMC (C) and HBdSF (D) cells after mock-knockdown or WISP1-knockdown were assessed by immunoblot assays. The mRNA levels of the WISP1, α-SMA, IL-6, GDF15, and CXCL5 after knockdown of WISP1 in the HBdSMC (E) and HBdSF (F) cells, as indicated, were determined by RT-qPCR. Data are presented as target genes/β-actin of mock-knockdown cells relative to WISP1-knockdown cells. ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Reverse Transcription Polymerase Chain Reaction, Knockdown, Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1-knockdown on the cell proliferation, migration, and contraction in bladder stroma and cancer cells. (A) The ability of cell proliferation of mock-knockdown HBdSMC (HBdSMC_shCOL) and WISPI-knockdown (HBdSMC_shWISP1) cells was assessed by EdU proliferation assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. (B) The ability of cell contraction of the mock-knockdown HBdSF (HBdSF_shCOL) and the WISP1-knockdown HBdSF (HBdSF_shWISP1) cells was measured by the collage contraction assays. Data are presented as gel area of the times as indicated in relation to the 0 h. (C) The ability of the migration of T24 cells after being treated with the supernatant from HBdSMC_shCOL and HBdSMC_shWISP1 cells, respectively, was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Knockdown, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1 on the cell proliferation and invasion in bladder cancer cells. (A) Cell proliferation of HT1376 and T24 cells after treatments of recombinant human WISP1 was assessed by CyQUANT cell proliferation assay. Data are presented as the mean percentage of the rhWISP1-treated cells relative to vehicle-treated cells. Expressions of total WISP1, WISP1v1, or WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the HT1376 cells were assessed by RT-qPCR (B), RT-PCR (C), and ELISA (D) assays. The abilities of the proliferation of HT-DNA, HT-WISP1v1 (E), and HT-WISP1v2 (F) cells were measured by Ki67 flow cytometry assays. Data are presented as the mean percentage of the Ki67-positive cells relative to HT-DNA cells. The invasion ability of HT1376 cells after ectopic overexpression of WISP1v1 (G) and WISP1v2 (H) was determined by in vitro Matrigel invasion assays (± SE; n = 4). Data are presented as the mean percentage of the invasion cells relative to HT-DNA cells. * p < 0.05; ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Recombinant, CyQUANT Assay, Proliferation Assay, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Over Expression, In Vitro

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Determination of the molecular mechanism of WISP1v1 on the cell invasion in the bladder cancer cells. (A) The protein levels of NDRG1, KAI1, and Maspin of HT-DNA and HT-WISP1v1 cells were determined by immunoblot assays. (B) The luciferase activity of the NDRG1 reporter vector was shown after transiently overexpressing various amounts of WISP1v1 expression vectors in HT1376 and T24 cells, as indicated. (C) The luciferase activity of KAI1 and Maspin reporter vectors was shown as indicated after transiently overexpressing various dosages of WISP1v1 expression vectors in HT1376 cells. (D) The luciferase activity of NDRG1, KAI1, and Maspin reporter vectors, respectively, after transiently overexpressed pcDNA, WISP1v1, and WISP1v2 expression vectors as indicated in HT1376 cells. (E) The mRNA levels of the E-cadherin, N-cadherin, Slug, Snail, and Vimentin were determined by RT-qPCR. Data are presented as target genes/β-actin of HT-WISP1v1 cells relative to HT-DNA cells. The mRNA levels of the WISP1 (F), NDRG1, KAI1, Maspin, and IL-6 (G) after transient overexpression of WISP1v1 in T24 cells were determined by RT-qPCR. Data are presented as target genes/β-actin of T24-WISP1v1 cells relative to T24-DNA cells. (H) The invasion ability of T24 cells after the ectopic overexpression of WISP1v1 was determined by in vitro Matrigel invasion assays (± SE; n = 4). The ability of the migration (I) and quantification analysis (I) of T24 cells after being treated with the supernatant from 293T-DNA and 293T-WISP1v1 cells was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. * p < 0.05, ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Western Blot, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Quantitative RT-PCR, Over Expression, In Vitro, Migration

Journal: bioRxiv

Article Title: WISP1 drives a mechanically active immune modulatory and proliferative cardiac myofibroblast state

doi: 10.64898/2026.02.17.706476

Figure Lengend Snippet: (A) Wisp1 mRNA expression in primary adult mouse cardiac fibroblasts following TGFβ1 (10 ng/mL) treatment for 24–72 h. (B) Representative immunofluorescence images of α-SMA (green), vimentin (red), and DAPI (blue) after 72 h treatment with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (C, D) Representative Western blot (C) and densitometric quantification (D) of α-SMA normalized to GAPDH. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05. Data are mean ± SEM from ≥3 independent isolations. Statistical analysis by one-way ANOVA with Tukey’s post hoc test; *p < 0.05.

Article Snippet: For treatments, CFs were seeded in 12- or 24-well plates in DMEM + 10% FBS and treated for 72 h with recombinant human TGFβ1 (10 ng/mL; R&D Systems, 7754-BH) and/or recombinant murine WISP1 (500 ng/mL; R&D Systems, 1680-WS-050).

Techniques: Expressing, Immunofluorescence, Western Blot

Journal: bioRxiv

Article Title: WISP1 drives a mechanically active immune modulatory and proliferative cardiac myofibroblast state

doi: 10.64898/2026.02.17.706476

Figure Lengend Snippet: Primary adult mouse cardiac fibroblasts were embedded in floating collagen gels and treated for 24 h with vehicle (Ctrl), WISP1 (500 ng/mL), TGFβ1 (10 ng/mL), or WISP1 + TGFβ1. (A) Representative gel images at 0 h and 24 h. (B) Quantification of gel area (% contraction; n = 4 isolations, 2 males & 2 females). (C–D) Confluent fibroblast monolayers were scratched and imaged every 2 hours for 24 hours using the same treatment conditions. (C) Representative bright-field images at time point 0 and 14. (D) Quantification of wound closure (% of gap closed relative to time 0; n = 8 isolations, 4 male & 4 female). Data are mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons; *p ≤ 0.05.

Article Snippet: For treatments, CFs were seeded in 12- or 24-well plates in DMEM + 10% FBS and treated for 72 h with recombinant human TGFβ1 (10 ng/mL; R&D Systems, 7754-BH) and/or recombinant murine WISP1 (500 ng/mL; R&D Systems, 1680-WS-050).

Techniques: