Therapeutic role of 2-stearoxyphenethyl phosphocholine targeting microtubule dynamics and Wnt/β-catenin/EMT signaling in human colorectal cancer cells

Author:

Park Sang-Eun1,Chung Kyung-Sook1,Kim Soo-Yeon1,Lee Jeong-Hun1,Hassan Ahmed H.E.1,Lee Yong Sup1,Lee Jae Yeol1,Lee Kyung -Tae1

Affiliation:

1. Kyung Hee University

Abstract

AbstractThe inhibition of cell death, perturbation of microtubule dynamics, and acceleration of Wnt/β-catenin/epithelial-mesenchymal transition (EMT) signaling are fundamental processes in the progression and metastasis of colorectal cancer (CRC). To explore the role of 2-stearoxyphenethyl phosphocholine (stPEPC), an alkylphospholipid-based compound, in CRC, we conducted an MTT assay, cell cycle analysis, western blot analysis, immunoprecipitation, immunofluorescence staining, Annexin V/propidium iodide double staining, small interfering RNA gene silencing, a wound-healing assay, an invasion assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay in the human CRC cell lines HT29 and HCT116. stPEPC showed anti-proliferative properties and mitotic cell accumulation via upregulated phosphorylation of BUBR1 and an association between mitotic arrest deficiency 2 (MAD2) and cell division cycle protein 20 homolog (CDC20). These results suggest that activation of the mitotic checkpoint complex and tubulin polymerization occurred, resulting in mitotic catastrophe in HT29 and HCT116 cells. In addition, stPEPC attenuated cell migration and invasion by regulating proteins mediated by EMT, such as E-cadherin and occludin. stPEPC downregulated the protein expression of Wnt3a and phosphorylation of low-density lipoprotein receptor-related protein 6 (LRP6), glycogen synthase kinase 3β (GSK3β), and β-catenin as well as their target genes, including cMyc and cyclin D1, in CRC cells. stPEPC may be useful for developing new drugs to treat human CRC.

Publisher

Research Square Platform LLC

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