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Enrichment of Cancer Stem-Like Cells by the Induction of Epithelial-Mesenchymal Transition Using Lentiviral Vector Carrying E-Cadherin Shrna in Ht29 Cell Line Publisher Pubmed



Saltanatpour Z1, 2 ; Johari B3 ; Alizadeh A4 ; Lotfinia M5 ; Majidzadeha K2 ; Nikbin B1 ; Kadivar M6
Authors
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Authors Affiliations
  1. 1. Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Genetics Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran
  3. 3. Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
  4. 4. Department of Tissue Engineering and Applied Cell Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran
  5. 5. Core Research Lab, Kashan University of Medical Sciences, Kashan, Iran
  6. 6. Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran

Source: Journal of Cellular Physiology Published:2019


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.
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