Rapid Adhesion and Spread of Non-adherent Colon Cancer Colo201 Cells Induced by the Protein Kinase Inhibitors, K252a and KT5720 and Suppression of the Adhesion by the Immunosuppressants FK506 and Cyclosporin A

Cell Structure and Function

Koshiro Hioki³  Shintaro Suzuki¹  Rikio Tokunaga²  Takashi Mohri³  Isamu Kameshita⁴  Shigeru Takatani² 

[1] Institute for Developmental Research, Aichi Human Service Center;Department of Hygiene, Kansai Medical University;Second Department of Surgery, Kansai Medical University;Department of Biochemistry, Asahikawa Medical School

关键词: colon cancer cells;    cell adhesion;    cell spreading;    K252a;    KT5720 and FK506;   

DOI  :  10.1247/csf.23.255

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(38)Cited-By(8)We examined alterations in cell morphology and expression of adhesion molecules in response to a general protein kinase inhibitor K252a treatment of non-adherent colon adenocarcinoma Colo201 cells. K252a induced rapid cell adhesion and spreading with concomitant formation of actin stress fibers. A protein kinase A inhibitor KT5720 also induced cell adhesion, but the rate of spread was slower than that seen with K252a. These adhesions were mediated by integrin molecules since cell adhesion required Mg2+, Mn2+ or Ca2+, and was inhibited by monoclonal antibodies for integrins α2 and β1. Indirect immunofluorescence microscopic observations revealed that integrin α2 and β1 molecules in K252a-treated cells were concentrated at sites of focal adhesion, but expressions of integrin molecules were not modulated. Tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin increased during K252a- or KT5720-induced cell adhesion. Immunosuppressants FK506 and cyclosporin A suppressed the K252a-induced cell adhesion and abolished tyrosine phosphorylation of cellular proteins including FAK and paxillin. Furthermore, W7 and calmidazolium, inhibitors of calmodulin, also inhibited the cell adhesion. Based on findings that FK506 and cyclosporin A are inhibitors of the calcium calmodulin-dependent protein phosphatase, calcineurin, this phosphatase may regulate integrin-dependent cell adhesion and spread of Colo201 cells. This Colo201 cell model provides a pertinent system for studying molecules involved in signal transduction pathways and can shed light on mechanisms of metastasis and invasion of colon carcinoma cells.

【 授权许可】

Unknown