Roghayeh Pourbagher1,2, Hossein Ghorbani3, Haleh Akhavan-Niaki2, Seyed Gholam Ali Jorsaraei4, Sadegh Fattahi2, Sahar Ghooran3, Zeinab Abedian1,5, Masoumeh Ghasemi2, Fatemeh Saeedi1,2, Negar Jafari1,2, Behnam Kalali6, Amrollah Mostafazadeh2. 1. Student Research Committee, Babol University of Medical Sciences, Babol, Iran. 2. Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran. 3. Department of Pathology, Rohani Hospital, Babol University of Medical Sciences, Babol, Iran. 4. Fatemeh Zahra Infertility and Reproductive Health Research Centre, Health Research Institute, Babol University of Medical Sciences, Babol, Iran. 5. Dental Materials Research Center, Dental Faculty, Babol University of Medical Sciences, Babol, Iran. 6. Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, Munich, Germany.
Abstract
BACKGROUND: Stem cell differentiation therapy is a promising strategy in cancer treatment. we show that protein cocktail prepared from serum starved fibroblasts has therapeutic potential based on this strategy. METHODS: The condition medium was prepared from foreskin isolated fibroblasts and analyzed by Liquid chromatography electrospray ionization mass spectrometry-mass spectrometry (LC-ESI-MS/MS). LA7 mammary gland cancer stem cells originated tumors were induced in Sprague Dawley rats. The rats treated subcutaneously with DMEM (group A), condition medium (group B), or normal saline (group C) once daily for 7 days. Then the tumors were removed and divided into the two parts, one part was used to quantify gene expression by stem-loop RT-qPCR assay and the other part was used for Hematoxylin & Eosin (H & E), Giemsa, and immunohistochemistry (IHC) staining. RESULTS: All induced tumors appeared as sarcomatoid carcinoma (SC). Immunohistochemistry staining confirmed this conclusion by recognizing the tumor as Ki67+, cytokeratin+, vimentine+, and estrogen receptor negative SC. RT-qPCR analysis revealed that Oct4-, Sox-2, Nanog- gene expression was much reduced in the condition medium treated tumors versus proper controls (p< 0.05). Tissue necrosis was more prevalent in this group while tumors volume was diminished almost by 40%. The LC-ESI-MS/MS analysis unrevealed the stemness reducing and the cell death inducing proteins such as, pigment epithelium-derived factor (PEDF), insulin like growth factor binding protein-5 (IGFBP-5) and -7 (IGFBP-7) in the condition medium. CONCLUSION: This study showed that the substances released from starved human fibroblasts were able to down-regulate the stemness-related genes and induce necrosis in LA7 derived tumors.
BACKGROUND: Stem cell differentiation therapy is a promising strategy in cancer treatment. we show that protein cocktail prepared from serum starved fibroblasts has therapeutic potential based on this strategy. METHODS: The condition medium was prepared from foreskin isolated fibroblasts and analyzed by Liquid chromatography electrospray ionization mass spectrometry-mass spectrometry (LC-ESI-MS/MS). LA7 mammary gland cancer stem cells originated tumors were induced in Sprague Dawley rats. The rats treated subcutaneously with DMEM (group A), condition medium (group B), or normal saline (group C) once daily for 7 days. Then the tumors were removed and divided into the two parts, one part was used to quantify gene expression by stem-loop RT-qPCR assay and the other part was used for Hematoxylin & Eosin (H & E), Giemsa, and immunohistochemistry (IHC) staining. RESULTS: All induced tumors appeared as sarcomatoid carcinoma (SC). Immunohistochemistry staining confirmed this conclusion by recognizing the tumor as Ki67+, cytokeratin+, vimentine+, and estrogen receptor negative SC. RT-qPCR analysis revealed that Oct4-, Sox-2, Nanog- gene expression was much reduced in the condition medium treated tumors versus proper controls (p< 0.05). Tissue necrosis was more prevalent in this group while tumors volume was diminished almost by 40%. The LC-ESI-MS/MS analysis unrevealed the stemness reducing and the cell death inducing proteins such as, pigment epithelium-derived factor (PEDF), insulin like growth factor binding protein-5 (IGFBP-5) and -7 (IGFBP-7) in the condition medium. CONCLUSION: This study showed that the substances released from starved human fibroblasts were able to down-regulate the stemness-related genes and induce necrosis in LA7 derived tumors.
Entities:
Keywords:
Breast cancer; Cancer Stem cells; Cell differentiation; Fibroblasts; Gene expression
Authors: Xinyu Tian; Bo Huang; Xiao-Peng Zhang; Mingyang Lu; Feng Liu; José N Onuchic; Wei Wang Journal: Proc Natl Acad Sci U S A Date: 2017-05-08 Impact factor: 11.205
Authors: I Zucchi; S Sanzone; S Astigiano; P Pelucchi; M Scotti; V Valsecchi; O Barbieri; G Bertoli; A Albertini; R A Reinbold; R Dulbecco Journal: Proc Natl Acad Sci U S A Date: 2007-06-12 Impact factor: 11.205