Increased expression of p21-activated kinase 4 in adenomyosis and its regulation of matrix metalloproteinase-2 and -9 in endometrial cells

Fertil Steril. 2015 Apr;103(4):1089-1097.e2. doi: 10.1016/j.fertnstert.2014.12.124. Epub 2015 Jan 27.

Abstract

Objective: To investigate the expression of p21-activated kinase 4 (Pak4) in both adenomyotic foci and the eutopic endometrium of women with adenomyosis, and whether the activities of matrix metalloproteinases (MMPs)-2 and -9 are regulated by Pak4 in endometrial cells.

Design: Experimental study using human samples and cell lines.

Setting: University hospital.

Patient(s): Thirty-nine patients with histologic evidence of adenomyosis, and 34 patients with carcinoma in situ of the uterine cervix without adenomyosis or endometriosis.

Intervention(s): Immunohistochemistry, zymography after transfection with Pak4 small interfering RNA (siRNA), and western blot analyses after nuclear factor kappa-B (NF-кB) inhibitor treatment.

Main outcome measure(s): The Pak4 immunoreactivity of women with vs. without adenomyosis was compared semiquantitatively. The activities of MMP-2 and -9 were analyzed in eutopic endometrial stromal cells and Ishikawa cells after transfection with Pak4 siRNA. The Pak4 expression was evaluated in endometrial cells after treatment with NF-кB inhibitor.

Result(s): Pak4 immunoreactivity was increased in adenomyotic foci and in the eutopic endometrium of women with adenomyosis. Transfection of endometrial cells with Pak4 siRNA led to significant decreases of MMP-2 and -9 activities. In vitro treatment of endometrial cells with tumor necrosis factor-alpha caused a significant increase of NF-кB activation and Pak4 expression, which was obviously decreased by the NF-кB inhibitor pyrrolidinedithiocarbamate.

Conclusion(s): Our results suggest that Pak4 is regulated by NF-кB and that increased Pak4 expression can lead to development of adenomyosis by enhancing the invasiveness of endometrial cells through regulation of MMP-2 and -9 activities.

Keywords: P21-activated kinase 4 (Pak4); adenomyosis; endometrium; matrix metalloproteinase (MMP); nuclear factor kappa-B (NF-кB).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenomyosis / genetics
  • Adenomyosis / metabolism*
  • Adenomyosis / pathology
  • Adult
  • Cell Movement / drug effects
  • Cell Movement / genetics
  • Cells, Cultured
  • Endometrium / metabolism*
  • Endometrium / pathology
  • Enzyme Activation / drug effects
  • Female
  • Humans
  • Immunohistochemistry
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinase 9 / metabolism*
  • RNA, Small Interfering / pharmacology
  • Up-Regulation / genetics
  • p21-Activated Kinases / genetics
  • p21-Activated Kinases / metabolism*
  • p21-Activated Kinases / physiology*

Substances

  • RNA, Small Interfering
  • PAK4 protein, human
  • p21-Activated Kinases
  • MMP2 protein, human
  • Matrix Metalloproteinase 2
  • MMP9 protein, human
  • Matrix Metalloproteinase 9