Enrichment of cancer stem‐like cells by the induction of epithelial‐mesenchymal transition using lentiviral vector carrying E‐cadherin shRNA in HT29 cell line. Issue 12 (20th May 2019)

• Record Type: Journal Article
• Authors: Saltanatpour, Zohreh; Johari, Behrooz; Alizadeh, Akram; Lotfinia, Majid; Majidzadeh‐A, Keivan; Nikbin, Behrooz; Kadivar, Mehdi
• Is Part Of: Journal of cellular physiology. Volume 234:Issue 12(2019:Dec.)
• Page Start: 22935
• Page End: 22946
• Publication Date: 2019-05-20
• Subjects: cancer stem cells (CSCs) -- cancer therapy. -- E‐cadherin -- epithelial‐mesenchymal transition (EMT) -- lentiviral vector; Physiology -- Periodicals; Cell physiology -- Periodicals; 571.6
• DOI: 10.1002/jcp.28855
• Languages: English
• ISSNs: 0021-9541

Abstract

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. 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.