Enrichment of cancer stem-like cells by the induction of epithelial-mesenchymal transition using lentiviral vector carrying E-cadherin shRNA in HT29 cell line

Saltanatpour, Z. and Johari, B. and Alizadeh, A. and Lotfinia, M. and Majidzadeh-A, K. and Nikbin, B. and Kadivar, M. (2019) Enrichment of cancer stem-like cells by the induction of epithelial-mesenchymal transition using lentiviral vector carrying E-cadherin shRNA in HT29 cell line. Journal of Cellular Physiology, 234 (12). pp. 22935-22946.

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DOI: UNSPECIFIED

Abstract

A better understanding of cancer stem cells (CSCs) may facilitate the prevention and treatment of cancers. Epithelial-mesenchymal transition (EMT) is a process activated during invasion and metastasis of tumors. EMT induction in normal and tumor cells makes them more resistant to chemotherapy. E-cadherin is a membrane protein and plays a role in tumor invasion, metastasis, and prognosis. Downregulation of E-cadherin is a hallmark of EMT. Here, we created a model of cancer stem-like cells enrichment via EMT induction using E-cadherin downregulation in HT29 cell line using a lentiviral vector carrying shRNA. We aimed to evaluate cancer and anti-CSC chemotherapeutics screening. The markers of EMT and CSCs were assessed and compared with control cells using flow cytometry, real-time PCR, immunocytochemistry, western blot, migration assay, invasion assay, and colony formation assay. The transduced cells showed a mesenchymal morphology. High levels of EMT-related proteins were also expressed. These results confirmed that the transduced cells underwent EMT. In addition, we observed an increased population of E-cadherin-downregulated HT29 cell line among the cells expressing colon CSC markers (CD133+ and CD44+) after EMT induction. E-cadherin-downregulated cells were morphologically like mesenchymal cells, and the number of CD133+- and CD44+-cells (CSC-like cells) increased. These cells can be used as stable models to study cancer cells and screening of antitumor therapeutics. © 2019 Wiley Periodicals, Inc.

Item Type: Article
Additional Information: cited By 1
Uncontrolled Keywords: CD133 antigen; Hermes antigen; lentivirus vector; short hairpin RNA; uvomorulin, Article; cancer stem cell; cell invasion assay; cell migration assay; colony formation; controlled study; down regulation; epithelial mesenchymal transition; flow cytometry; HT-29 cell line; human; immunocytochemistry; priority journal; real time polymerase chain reaction; Western blotting
Subjects: Pathology
Biochemistry, Genetics and Molecular Biology
Divisions: Faculty of Medicine > Basic Sciences > Department of Pathology& Histology
Depositing User: ART . editor
Date Deposited: 24 Dec 2019 12:25
Last Modified: 24 Dec 2019 12:25
URI: http://eprints.kaums.ac.ir/id/eprint/4805

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