SPAK mediates KCC3-enhanced cervical cancer tumorigenesis

Min Hsi Chiu, Hsiao Sheng Liu, Yi Hui Wu, Meng Ru Shen, Cheng Yang Chou

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)


Ste20-related proline/alanine-rich kinase (SPAK) plays a role in regulating many biological activities, and interacts with K-Cl co-transporter 3 (KCC3); however, the importance of SPAK for KCC3 function has not been demonstrated. Here, we investigated the role of SPAK in KCC3-regulated cell invasiveness and tumor formation. We show that induction of KCC3 expression triggers activation of the NF-κB and SPAK signaling cascades, leading to activation of p38 mitogen-activated protein kinase (MAPK) and matrix metalloproteinase-2 (MMP2). A small interference RNA-mediated reduction in SPAK protein levels suppressed the invasive ability and oncogenic potential of cervical cancer cells, and decreased tumor formation in mouse xenografts. A combination of experimental approaches, including RT-PCR and real-time RT-PCR, gelatin zymography, NF-κB luciferase activity and chromatin immunoprecipitation assays, showed that the induction of KCC3 over-expression increased MMP2 expression and augmented binding of NF-κB to its putative SPAK promoter binding site, suggesting that the SPAK/MMP2 axis is up-regulated by NF-κB. Pharmacological inhibition of NF-κB or MMP2 abrogated KCC3-triggered, SPAK-dependent cell invasiveness. Furthermore, p38 MAPK was identified as the upstream regulator of KCC3-dependent MMP2 activation. We conclude that SPAK may promote KCC3-mediated tumor aggressiveness via the NF-κB/p38 MAPK/MMP2 axis. Structured digital abstract KCC3 physically interacts with SPAK by anti-bait co-immunoprecipitation (view interaction) SPAK plays a role in regulating many biological activities and interacts with the KCC3. In this study, we show that KCC3-induced SPAK expression occurs through binding of the transcription factor NF-κB to the SPAK promoter, thereby promoting cell aggressiveness. We concluded that SPAK may promote the KCC3-mediated tumor aggressiveness via the NF-κB/p38 MAPK/MMP2 axis.

Original languageEnglish
Pages (from-to)2353-2365
Number of pages13
JournalFEBS Journal
Issue number10
Publication statusPublished - 2014 May

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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