A novel tumor suppressor role of myosin light chain kinase splice variants through downregulation of the TEAD4/CD44 axis

Yen-Ju Huang, Tsung-Chun Lee, Yu-Chen Pai, Been-Ren Lin, Jerrold R Turner, Linda Chia-Hui Yu

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Myosin light chain kinase (MLCK) regulates actinomyosin contraction. Two splice variants of long MLCK are expressed in epithelial cells and divergently regulate gut barrier functions; reduced MLCK levels in human colorectal cancers (CRC) with unclarified significance have been reported. CRC are solid tumors clonally sustained by stem cells highly expressing CD44 and CD133. The aim was to investigate the role of MLCK splice variants in CRC tumorigenesis. We found lower MLCK1/2 and higher CD44 expression in human CRC, but no change in CD133 or LGR5. Large-scale bioinformatics showed an inverse relationship between MYLK and CD44 in human sample gene datasets. A 3-fold increased tumor burden was observed in MLCK(-/-) mice compared with wild-type (WT) mice in a chemical-induced CRC model. Primary tumorspheres derived from the MLCK(-/-) mice displayed larger sizes and higher CD44 transcript levels than those from the WT mice. Bioinformatics revealed binding of TEAD4 (a transcriptional enhancer factor family member in the Hippo pathway) to CD44 promoter, which was confirmed by luciferase reporter assay. Individually expressing MLCK1 and MLCK2 variants in the MLCK-knockout (KO) Caco-2 cells inhibited the nuclear localization of TEAD4 cofactors, VGLL3 and YAP1, respectively, and both variants reduced the CD44 transcription. Accelerated cell cycle transit was observed in the MLCK-KO cells, whereby expression of MLCK1/2 variants counterbalanced the cell hyperproliferation. In conclusion, MLCK1/2 variants are novel tumor suppressors by downregulating the TEAD4/CD44 axis via reducing nuclear translocation of distinct transcriptional coactivators. The reduction of epithelial MLCKs, especially isoform 2, may drive cancer stemness and tumorigenesis.

Original languageEnglish
Pages (from-to)961-974
Number of pages14
JournalCarcinogenesis
Volume42
Issue number7
DOIs
Publication statusPublished - Jul 16 2021
Externally publishedYes

Keywords

  • Adaptor Proteins, Signal Transducing/genetics
  • Alternative Splicing
  • Animals
  • Apoptosis
  • Biomarkers, Tumor/genetics
  • Cell Cycle
  • Cell Movement
  • Cell Proliferation
  • Colonic Neoplasms/genetics
  • DNA-Binding Proteins/genetics
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Hyaluronan Receptors/genetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Muscle Proteins/genetics
  • Myosin-Light-Chain Kinase/genetics
  • Phosphorylation
  • Prognosis
  • Survival Rate
  • TEA Domain Transcription Factors
  • Transcription Factors/genetics
  • Tumor Cells, Cultured
  • YAP-Signaling Proteins

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