tβri-based cancer therapies and biomarkers  (Biotech AB)

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Exon 2 of TβRI was designed to be targeted by the mutant Cas9 enzyme. The mutant clone A9 contained a deletion of TβRI as detected by sequencing and the stop sequence formed by a frameshift mutation. b The mutant cell clones (A–E) and wild-type (WT) cells were examined by Cel-1 assay (Surveyor nuclease assay). c Mutant clones B5 and A9 and WT cells detected by RT-PCR. d , e Western blot and f qPCR were used to verify the TβRI-deficient cell lines. Results are shown as mean ± SEM, * p < 0.05, ** p < 0.01, Student’s t -test. g WT, A9, and HA - TβR1 reconstituted A9 cells were lysed and subjected to immunoblotting with antibodies TβRI, pSmad2, Smad2, and actin. h CAGA-luciferase reporter assay showing the TGFβ response of the indicated cell lines. Results are shown as mean ± SEM, * p < 0.05, *** p < 0.001, Student’s t -test.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Mutagenesis, Sequencing, Clone Assay, Nuclease Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Luciferase, Reporter Assay

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Gene-modified cell lines wild-type (WT), A9 (TβRI deficient), A9 HA-TβRI (A9 cells reconstituted with HA-TβRI), and A9 vector (empty vector as control) were subjected to immunoblotting with antibodies pSmad2, Smad2, TβRI, and actin. b CAGA-luciferase reporter assay showing the TGFβ response in the indicated cell lines. Results are shown as mean ± SEM, *** p < 0.001, Student’s t -test. c Expression of Smad7 and SERPINE1 in the indicated cell lines detected by qPCR. Results are shown as mean ± SEM, *** p < 0.001, Student’s t -test. d Microarray data showing the differentially expressed genes (DEGs) in the indicated cell lines with or without TGFβ treatment. e DEGs were annotated to the biological functions by DAVID Bioinformatics Resources. * p < 0.05, ** p < 0.01.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Modification, Plasmid Preparation, Control, Western Blot, Luciferase, Reporter Assay, Expressing, Microarray

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a – c Expression of THBS1 and ITGAV detected by western blotting in the PC3U, DU-145, and A549 cell lines with or without a knockout or knockdown of TβRI. d Immunofluorescent (IF) staining of THBS1 and pSmad2 in WT and A9 cells in a wound-healing assay with or without TGFβ treatment. Scale bar, 20 μm. e Invasion assay showing the invasive capacity of PC3U cells with or without knockout of TβRI or with or without TGFβ treatment for 48 h. Scale bar, 50 μm. f Western blotting showing the overexpression of THBS1 in A9 cells. g Invasion assay showing the invasive capacity of WT, A9, and A9 cells overexpressed THBS1. Scale bar, 50 μm. h The invading cells were quantified by OD value. Shown as mean ± SD, * p < 0.05. i IF staining of THBS1 and Paxillin in PC3U cells treated with TGFβ for 6 h. Scale bar, 20 μm. j IF staining of THBS1, Paxillin, and Phalloidin in PC3U cells treated with TGFβ for 6 h. Scale bar, 20 μm.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Expressing, Western Blot, Knock-Out, Knockdown, Staining, Wound Healing Assay, Invasion Assay, Over Expression

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Co-IF staining of HA, THBS1, and ITGAV in the wild-type (WT), A9 (TβRI deficient), A9 HA-TβRI (A9 cells reconstituted with HA-TβRI) cell lines with or without TGFβ treatment. Scale bar, 20 μm. b Co-IP was performed to detect the interaction of HA-TβRI with THBS1 and ITGAV. c The correlation of TβRI and THBS1 expression, TβRI and ITGAV expression in prostate cancer (TCGA database). d The interaction of THBS1 and ITGAV was detected in a wound-healing assay by in situ PLA. Scale bar, 50 μm. e Invasion assay showing that knockdown of THSB1 prevented the invasion capacity of PC3U cells treated with TGFβ. Scale bar, 50 μm. f The invading cells were quantified by OD value. Shown as mean ± SD, * p < 0.05. g Western blot showing the expression of THSB1 and ITGAV in PC3U cells with THSB1 knockdown.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Staining, Co-Immunoprecipitation Assay, Expressing, Wound Healing Assay, In Situ, Invasion Assay, Knockdown, Western Blot

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Wild-type (WT) PC3U and TβRI-deficient A9 cells were injected into the prostates of mice. Tumors and sentinel lymph nodes were obtained from the orthotopic xenograft mouse model after 4 weeks. b The tumor weight was quantified in both the WT ( n = 9) and A9 ( n = 9) groups. Results are shown as mean ± SEM, *** p < 0.001, one-way ANOVA. c The metastatic lymph nodes were quantified in both the WT ( n = 9) and A9 ( n = 6) groups. Results are shown as mean ± SEM, *** p < 0.001, one-way ANOVA. d Representative images of THBS1 staining in the WT or A9 tumors and lymph metastasis. Scale bar, 500 μm. Left: the black arrow shows the lymph vessels in the tumor-invasive front, and the red arrow shows tumor cells infiltrating into the adjacent muscles. Right: the black arrows show the remaining lymphatic cells in the sentinel lymph nodes. Scale bar, 100 μm. e Representative images of THBS1 staining in the normal adjacent prostate tissue, lower malignant prostatic adenocarcinoma (GS ≤ 6), and higher malignant prostatic adenocarcinoma (GS > 6). f THBS1 expression was examined on tissue microarray sections with 192 samples. Immunoreactivity (IR) was scored as negative (IR = 0), weak (IR = 1), moderate (IR = 2), or strong (IR = 3) **** p < 0.0001, Tukey’s multiple comparisons test. g A proteomic database was used to analyze the THBS1 protein level in the control adjacent prostate tissue ( n = 8), localized tumor ( n = 28), and bone metastasis ( n = 22). **** p < 0.0001, one-way ANOVA. h Representative images for the IF staining of THBS1 and panCK AE1/3 in the human primary and bone metastatic tumors. Scale bar, 50 μm.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Injection, Staining, Muscles, Expressing, Microarray, Control