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Melatonin inhibits the Migration of Colon Cancer RKO cells by Down-regulating Myosin Light Chain Kinase Expression through Cross-talk with p38 MAPK

  • Zou, Duo-Bing (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Wei, Xiao (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Hu, Ruo-Lei (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Yang, Xiao-Ping (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Zuo, Li (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Zhang, Su-Mei (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Zhu, Hua-Qing (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Zhou, Qing (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University) ;
  • Gui, Shu-Yu (Key Laboratory of Gene Research of Anhui Province) ;
  • Wang, Yuan (Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University)
  • Published : 2015.09.02

Abstract

Background: Melatonin, which is mainly produced by the pineal gland, has a good inhibitory effect on cell growth of multiple cancer types. However, the underlying molecular mechanisms of anti-tumor activity for colon cancer have not been fully elucidated. In this study, we investigated the effects of melatonin on migration in human colon cancer RKO cells and the potential molecular mechanisms. Materials and Methods: The viability of RKO cells was investigated by MTT assay after treatment with melatonin, SB203580 (p38 inhibitor) and phorbol 12-myristate 13-acetate (PMA, MAPK activator) alone or in combination for 48h. The effects of melatonin, and ML-7, a selective inhibitor of myosin light chain kinase (MLCK), and SB203580, and PMA on the migration of RKO cells were analyzed by in vitro scratch-wound assay. The relative mRNA levels of MLCK was assessed by real-time quantitative RT-PCR. Western blotting analysis was performed to examine the expression of MLCK, phosphorylation of myosin light chain (pMLC) and p38 (pp38). Results: The proliferation and migration of human colon cancer RKO cells were inhibited significantly after treatment with melatonin. The expression levels of MLCK and phosphorylation of MLC of RKO cells were reduced, and real-time quantitative RT-PCR showed that melatonin had significant effects on suppressing the expression of MLCK. Furthermore, the phosphorylation level of p38, which showed the same trend, was also reduced when cells were treated by melatonin. In addition, ML-7 (25umol/l) could down-regulate the phosphorylation of p38. Conclusions: Melatonin could inhibit the proliferation and migration of RKO cells, and further experiments confirmed that p38 MAPK plays an important role in regulating melatonin-induced migration inhibition through down-regulating the expression and activity of MLCK.

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