Identification of Fibulin‐1 as a Human Bone Marrow Stromal (HS‐5) Cell‐Derived Factor That Induces Human Prostate Cancer Cell Death
Autor: | Kelvin H. Lee, Leila H. Choe, Irene R. Marwa, Robert A. Sikes, Kornkamon Lertsuwan |
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Rok vydání: | 2017 |
Předmět: |
Male
0301 basic medicine Pathology medicine.medical_specialty Programmed cell death Stromal cell Cell Survival Urology Apoptosis Bone Neoplasms Biology Transforming Growth Factor beta1 03 medical and health sciences Prostate cancer 0302 clinical medicine Prostate Cell Line Tumor medicine Humans Cell Death Calcium-Binding Proteins Prostatic Neoplasms Bone metastasis Mesenchymal Stem Cells medicine.disease 030104 developmental biology medicine.anatomical_structure Oncology Culture Media Conditioned 030220 oncology & carcinogenesis Cancer cell Cancer research Bone marrow |
Zdroj: | The Prostate. 77:729-742 |
ISSN: | 1097-0045 0270-4137 |
DOI: | 10.1002/pros.23303 |
Popis: | BACKGROUND Previous studies showed that human bone marrow stromal HS-5 cells secreted unidentified factor(s) inducing PCa cell death. Herein, the HS-5-derived factor (HS-5 DF) was characterized and identified. METHODS Conditioned media from confluent HS-5 cells were collected and modified for biochemical characteristic testing of HS-5 DF. Cell survival was measured by apoptosis assay and live/dead assay. Fibulin-1 was identified from gel electrophoresis and mass spectrometry. The validation of Fibulin-1 as a HS-5 DF was done by immunoprecipitation (IP) and genetic knockdown by CRISPR/Cas9 system. RESULTS HS-5 DF was trypsin and heat sensitive, but pH stable. The tentative size of the factor fell between 30 kDa and 100 kDa. TGF-β1 treatment led to a suppression of HS-5 DF activity, a property consistent with bone metastasis in prostate cancer. Examination of TGF-β1 down regulated proteins led to identification of fibulin-1 as a candidate for the DF. IP of Fibulin-1 from HS-5 CM and CRISPR knockdown of Fibulin-1 showed a significant reduction of HS-5 CM-derived PCa cell death. These results strongly support a role for fibulin-1 in HS-5 bone marrow stromal cell induction of PCa cell death. CONCLUSION Our data indicate that Fibulin-1 functions as a HS-5 bone marrow stromal cell-derived factor inducing prostate cancer cell death. Prostate © 2016 Wiley Periodicals, Inc. |
Databáze: | OpenAIRE |
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