Lithium chloride induces mesenchymal‑to‑epithelial reverting transition in primary colon cancer cell cultures

International Journal of Oncology
Valeria CostabileMarina De Rosa

Abstract

Epithelial‑to‑mesenchymal transition (EMT) confers stem cell‑like phenotype and more motile properties to carcinoma cells. During EMT, the expression of E‑cadherin decreases, resulting in loss of cell‑cell adhesion and increased migration. Expression of Twist1 and other pleiotropic transcription factors, such as Snail, is known to activate EMT. We established primary colon cancer cell cultures from samples of operated patients and validated cultures by cytogenetic and molecular biology approaches. Western blot assay, quantitative real‑time PCR and immunofluorescence were performed to investigate the expression of E‑cadherin, vimentin, β‑catenin, cytokeratin‑20 and ‑18, Twist1, Snail, CD44, cyclooxygenase‑2 (COX2), Sox2, Oct4 and Nanog. Moreover, cell differentiation was induced by incubation with LiCl‑containing medium for 10 days. We observed that these primary colorectal cancer (CRC) cells lost expression of the E‑cadherin epithelial marker, which was instead expressed in cancer and normal colon mucosa of the same patient, while overexpressed vimentin (mesenchymal marker), Twist1, Snail (EMT markers) and COX2. Cytokeratin‑18 was expressed both in tissues and cell cultures. Expression of stem cell markers, such as CD44, Oct4 a...Continue Reading

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Citations

Nov 12, 2017·The Journal of Membrane Biology·Eric JakobssonLaura Pritchet
Jul 8, 2015·Oncology Reports·Marina De RosaPaolo Delrio
Apr 5, 2016·Asian Pacific Journal of Cancer Prevention : APJCP·Elsayed I SalimHager M Helmy
Jun 7, 2020·Cells·Takahiro DomotoToshinari Minamoto
May 23, 2019·Frontiers in Oncology·Weihao Ge, Eric Jakobsson
Feb 3, 2021·Cells·Melinda E SnitowPeter S Klein

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Methods Mentioned

BETA
ubiquitination
electrophoresis
PCR
protein assay

Software Mentioned

BLAST
Primer
ISIS

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