Downregulation of Stemness Genes and Induction of Necrosis in Rat LA۷ Cancer Stem Cells Induced Tumors Treated with Starved Fibroblasts Culture Supernatant

Publish Year: 1400
نوع سند: مقاله ژورنالی
زبان: English
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JR_RBMB-10-1_012

تاریخ نمایه سازی: 17 خرداد 1400

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). LA۷ 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 ۷ 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 Ki۶۷+, cytokeratin+, vimentine+, and estrogen receptor negative SC. RT-qPCR analysis revealed that Oct۴-, Sox-۲, Nanog- gene expression was much reduced in the condition medium treated tumors versus proper controls (p< ۰.۰۵). Tissue necrosis was more prevalent in this group while tumors volume was diminished almost by ۴۰%. 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-۵ (IGFBP-۵) and -۷ (IGFBP-۷) in the condition medium. Conclusions: This study showed that the substances released from starved human fibroblasts were able to down-regulate the stemness-related genes and induce necrosis in LA۷ derived tumors.

Authors

Roghayeh Pourbagher

Student Research Committee, Babol University of Medical Sciences, Babol, Iran & Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Hossein Ghorbani

Department of Pathology, Rohani Hospital, Babol University of Medical Sciences, Babol, Iran.

Haleh Akhavan-Niaki

Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Seyed Gholam Ali Jorsaraei

Fatemeh Zahra Infertility and Reproductive Health Research Centre, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Sadegh Fattahi

Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Sahar Ghooran

Department of Pathology, Rohani Hospital, Babol University of Medical Sciences, Babol, Iran.

Zeinab Abedian

Student Research Committee, Babol University of Medical Sciences, Babol, Iran & Dental Materials Research Center, Dental Faculty, Babol University of Medical Sciences, Babol, Iran.

Masoumeh Ghasemi

Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Fatemeh Saeedi

Student Research Committee, Babol University of Medical Sciences, Babol, Iran & Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Negar Jafari

Student Research Committee, Babol University of Medical Sciences, Babol, Iran & Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Behnam Kalali

Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, Munich, Germany.

Amrollah Mostafazadeh

Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

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