Overexpression of dnIKK in mesenchymal stem cells leads to increased migration and decreased invasion upon TNFα stimulation

Biochem Biophys Res Commun. 2013 Jun 28;436(2):265-70. doi: 10.1016/j.bbrc.2013.05.091. Epub 2013 Jun 4.

Abstract

IκB kinase 2 (IKK-2) mediates tumor necrosis-factor α (TNFα) induced invasion of human mesenchymal stem cell (hMSC) to sites of tissue injury. Suppressing IKK-2 activity leads to reduced expression of proteolytic enzymes and impaired invasive capacity. In order to further reveal mechanisms of hMSC recruitment, we here aimed to analyse the impact of IKK-2 on two-dimensional migration upon TNFα stimulation in contrast to three-dimensional invasion. An immortalized hMSC line (SCP-1) was transduced with a dominant-negative mutant of IκB kinase 2 (SCP-1 dnIKK). Migration was assessed using a linear-gradient chemotaxis chambers by time-lapse analysis. Invasive capacity through human extracellular matrix was analysed using transwell invasion assays. RT-PCR confirmed increased IKK-2 expression levels in SCP-1 dnIKK cells, while TNFα receptor I and II expression was not altered. Invasion upon TNFα stimulation was significantly reduced by 78% in SCP-1 dnIKK. In contrast, migration was significantly increased, represented by a 60% elevated forward migration index and a 2.1-fold higher mean dislocation of the center of mass towards TNFα. In conclusion, our data confirms the impact of IKK-2 in TNFα dependent hMSC recruitment. Interestingly, reducing IKK-2 function increases two-dimensional migration towards TNFα, while invasive capacity is impaired. These findings contribute to a deeper understanding of MSC's biological properties orchestrating the complex processes of stem cell recruitment and homing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Movement / drug effects*
  • Chemotaxis / drug effects
  • Gene Expression / drug effects
  • Humans
  • I-kappa B Kinase / genetics
  • I-kappa B Kinase / metabolism*
  • Immunohistochemistry
  • Lentivirus / genetics
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects*
  • Mesenchymal Stem Cells / metabolism
  • Receptors, Tumor Necrosis Factor / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor RelA / metabolism
  • Transduction, Genetic
  • Tumor Necrosis Factor-alpha / pharmacology*

Substances

  • Receptors, Tumor Necrosis Factor
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • I-kappa B Kinase