All-Trans Retinoic Acid Inhibits Human Colorectal Cancer Cells RKO Migration via Downregulating Myosin Light Chain Kinase Expression through MAPK Signaling Pathway

ABSTRACT All-trans-retinoic acid (ATRA) inhibits the invasive and metastatic potentials of various cancer cells. However, the underlying mechanism is unclear. Here, we demonstrate that ATRA inhibited colorectal cancer cells RKO (human colon adenocarcinoma cell) migration by downregulating cell movement and increasing cell adhesion. ATRA inhibited the expression and activation of myosin light chain kinase (MLCK) in RKO cells, while the expression level of MLC phosphatase (MLCP) had no change in RKO cells treated with or without ATRA. The expression and activity of MLC was also inhibited in RKO cells exposed to ATRA. Intriguingly, ATRA increased the expression of occludin messenger RNA (mRNA) and protein and its localization on cell membrane. However, ATRA did not change the expression of zonula occludens 1 (ZO-1), but increased the accumulation of ZO-1 on RKO cells membrane. ML-7, an inhibitor of MLCK, significantly inhibited RKO cell migration. Furthermore, knockdown of endogenous MLCK expression inhibited RKO migration. Mechanistically, we showed that MAPK-specific inhibitor PD98059 enhanced the inhibitory effect of ATRA on RKO migration. In contrast, phorbol 12-myristate 13-acetate (PMA) attenuated the effects of ATRA in RKO cells. Moreover, knocking down endogenous extracellular signal-regulated kinase (ERK) expression inhibited MLCK expression in the RKO cells. In conclusion, ATRA inhibits RKO migration by reducing MLCK expression via extracellular signal-regulated kinase 1/Mitogen-activated protein kinase (ERK1/MAPK) signaling pathway.

[1]  Jiaoti Huang,et al.  All-trans retinoic acids induce differentiation and sensitize a radioresistant breast cancer cells to chemotherapy , 2016, BMC Complementary and Alternative Medicine.

[2]  G. Tonini,et al.  Pseudocirrhosis after anti-EGFR-based neoadjuvant therapy for hepatic metastasis from colon cancer: a different point of view. , 2014, Clinical colorectal cancer.

[3]  S. Garattini,et al.  Retinoids and breast cancer: from basic studies to the clinic and back again. , 2014, Cancer treatment reviews.

[4]  D. Early,et al.  Patient attitudes and issues in colon cancer screening. , 2014, Journal of the National Comprehensive Cancer Network : JNCCN.

[5]  S. Ichinose,et al.  Myosin Light Chain Kinase Expression Induced via Tumor Necrosis Factor Receptor 2 Signaling in the Epithelial Cells Regulates the Development of Colitis-Associated Carcinogenesis , 2014, PloS one.

[6]  S. Yuan,et al.  Myosin Light Chain Kinase Signaling in Endothelial Barrier Dysfunction , 2013, Medicinal research reviews.

[7]  Jie Gao,et al.  All-Trans-Retinoic Acid Modulates ICAM-1 N-Glycan Composition by Influencing GnT-III Levels and Inhibits Cell Adhesion and Trans-Endothelial Migration , 2012, PloS one.

[8]  W. Hauswirth,et al.  ERK1/2 activation is a therapeutic target in age-related macular degeneration , 2012, Proceedings of the National Academy of Sciences.

[9]  J. Turner,et al.  Myosin light chain kinase: pulling the strings of epithelial tight junction function , 2012, Annals of the New York Academy of Sciences.

[10]  Le Shen.  Tight junctions on the move: molecular mechanisms for epithelial barrier regulation , 2012, Annals of the New York Academy of Sciences.

[11]  P. Dollé,et al.  Retinoic acid signalling during development , 2012, Development.

[12]  Siddikuzzaman,et al.  All Trans Retinoic Acid and Cancer , 2011, Immunopharmacology and immunotoxicology.

[13]  N. Kemeny,et al.  Metastatic Colorectal Cancer: From Improved Survival to Potential Cure , 2010, Oncology.

[14]  C. Weber,et al.  MLCK-dependent exchange and actin binding region-dependent anchoring of ZO-1 regulate tight junction barrier function , 2010, Proceedings of the National Academy of Sciences.

[15]  D. Cui,et al.  Inhibitory effects of retinoic acid on invasiveness of human thyroid carcinoma cell lines in vitro , 2009, Journal of endocrinological investigation.

[16]  Paula D. Bos,et al.  Metastasis: from dissemination to organ-specific colonization , 2009, Nature Reviews Cancer.

[17]  J. Strong,et al.  Stimulation of Ebola virus production from persistent infection through activation of the Ras/MAPK pathway , 2008, Proceedings of the National Academy of Sciences.

[18]  Jenny Y. Bang,et al.  Antitumor effect of all-trans retinoic acid-encapsulated nanoparticles of methoxy poly(ethylene glycol)-conjugated chitosan against CT-26 colon carcinoma in vitro. , 2008, Journal of pharmaceutical sciences.

[19]  J. Miyoshi,et al.  Structural and functional associations of apical junctions with cytoskeleton. , 2008, Biochimica et biophysica acta.

[20]  A. Malik,et al.  Regulation of Endothelial Junctional Permeability , 2008, Annals of the New York Academy of Sciences.

[21]  E. Park,et al.  Retinol Inhibits the Invasion of Retinoic Acid–Resistant Colon Cancer Cells In Vitro and Decreases Matrix Metalloproteinase mRNA, Protein, and Activity Levels , 2007, Nutrition and cancer.

[22]  D. Rohde,et al.  All-Trans Retinoic Acid and Interferon-α for Treatment of Human Renal Cell Carcinoma Multicellular Tumor Spheroids , 2004, Urologia Internationalis.

[23]  D. Hartshorne,et al.  Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts , 2004, The Journal of cell biology.

[24]  Donna J. Webb,et al.  FAK–Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly , 2004, Nature Cell Biology.

[25]  E. Dmitrovsky,et al.  Retinoids in cancer therapy and chemoprevention: promise meets resistance , 2003, Oncogene.

[26]  Qiao Wu,et al.  Effects of retinoic acid on metastasis and its related proteins in gastric cancer cells in vivo and in vitro. , 2002, Acta pharmacologica Sinica.

[27]  C. Sigman,et al.  Retinoids in chemoprevention and differentiation therapy. , 2000, Carcinogenesis.

[28]  P. Verkade,et al.  Tight junctions are membrane microdomains. , 2000, Journal of cell science.

[29]  S. L. Gonias,et al.  Myosin Light Chain Kinase Functions Downstream of Ras/ERK to Promote Migration of Urokinase-Type Plasminogen Activator-Stimulated Cells in an Integrin-Selective Manner , 1999, The Journal of cell biology.

[30]  David A. Cheresh,et al.  Regulation of Cell Motility by Mitogen-activated Protein Kinase , 1997, The Journal of cell biology.

[31]  A. Malik,et al.  Regulation of vascular endothelial barrier function. , 1994, The American journal of physiology.

[32]  Yang Bi,et al.  All-trans retinoic acid inhibits proliferation, migration, invasion and induces differentiation of hepa1-6 cells through reversing EMT in vitro. , 2016, International journal of oncology.

[33]  Yuan Wang,et al.  Butyrate suppresses proliferation and migration of RKO colon cancer cells though regulating endocan expression by MAPK signaling pathway. , 2013, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[34]  D. Ye,et al.  TNF-a Modulation of Intestinal Epithelial Tight Junction Barrier Is Regulated by ERK1/2 Activation of Elk-1 , 2013 .