Motor protein-dependent membrane trafficking of KCl cotransporter-4 is important for cancer cell invasion

Yih Fung Chen, Cheng-Yang Chou, Robert J. Wilkins, J. Clive Ellory, David B. Mount, Meng-Ru Shen

Research output: Contribution to journalArticle

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Abstract

The KCl cotransporter (KCC) is a major determinant of osmotic homeostasis and plays an emerging role in tumor biology. This study stresses the important role of KCC4 in tumor malignant behavior. Real-time reverse transcription-PCR on samples collected by laser microdissection and immunofluorescent stainings with different KCC isoform antibodies indicate that KCC4 is abundant in metastatic cervical and ovarian cancer tissues. Insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) stimulate KCC4 recruitment from a presumably inactive cytoplasmic pool of endoplasmic reticulum and Golgi to plasma membrane along actin cytoskeleton that is significantly inhibited by LY294002 and wortmannin. Throughout the trafficking process, KCC4 is incorporated into lipid rafts that function as a platform for the association between KCC4 and myosin Va, an actin-dependent motor protein. KCC4 and ezrin, a membrane cytoskeleton linker, colocalize at lamellipodia of migratory cancer cells. Interference with KCC activity by either an inhibitor or a dominant-negative loss-of-function mutant profoundly suppressed the IGF-I-induced membrane trafficking of KCC4 and the structural interaction between KCC4 and ezrin near the cell surface. Endogenous cancer cell invasiveness was significantly attenuated by small interfering RNA targeting KCC4, and the residual invasiveness was much less sensitive to IGF-I or EGF stimulation. In the metastatic cancer tissues, KCC4 colocalizes with IGF-I or EGF, indicating a likely in vivo stimulation of KCC4 function by growth factors. Thus, blockade of KCC4 trafficking and surface expression may provide a potential target for the prevention of IGF-I- or EGF-dependent cancer spread.

Original languageEnglish
Pages (from-to)8585-8593
Number of pages9
JournalCancer Research
Volume69
Issue number22
DOIs
Publication statusPublished - 2009 Nov 15

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Insulin-Like Growth Factor I
Membrane Proteins
Epidermal Growth Factor
Neoplasms
Microdissection
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Pseudopodia
Membranes
Myosins
Cytoskeleton
Actin Cytoskeleton
Uterine Cervical Neoplasms
Endoplasmic Reticulum
Ovarian Neoplasms
Small Interfering RNA
Reverse Transcription
potassium-chloride symporters
Actins
Intercellular Signaling Peptides and Proteins
Protein Isoforms

All Science Journal Classification (ASJC) codes

  • Cancer Research
  • Oncology

Cite this

Chen, Yih Fung ; Chou, Cheng-Yang ; Wilkins, Robert J. ; Ellory, J. Clive ; Mount, David B. ; Shen, Meng-Ru. / Motor protein-dependent membrane trafficking of KCl cotransporter-4 is important for cancer cell invasion. In: Cancer Research. 2009 ; Vol. 69, No. 22. pp. 8585-8593.
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abstract = "The KCl cotransporter (KCC) is a major determinant of osmotic homeostasis and plays an emerging role in tumor biology. This study stresses the important role of KCC4 in tumor malignant behavior. Real-time reverse transcription-PCR on samples collected by laser microdissection and immunofluorescent stainings with different KCC isoform antibodies indicate that KCC4 is abundant in metastatic cervical and ovarian cancer tissues. Insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) stimulate KCC4 recruitment from a presumably inactive cytoplasmic pool of endoplasmic reticulum and Golgi to plasma membrane along actin cytoskeleton that is significantly inhibited by LY294002 and wortmannin. Throughout the trafficking process, KCC4 is incorporated into lipid rafts that function as a platform for the association between KCC4 and myosin Va, an actin-dependent motor protein. KCC4 and ezrin, a membrane cytoskeleton linker, colocalize at lamellipodia of migratory cancer cells. Interference with KCC activity by either an inhibitor or a dominant-negative loss-of-function mutant profoundly suppressed the IGF-I-induced membrane trafficking of KCC4 and the structural interaction between KCC4 and ezrin near the cell surface. Endogenous cancer cell invasiveness was significantly attenuated by small interfering RNA targeting KCC4, and the residual invasiveness was much less sensitive to IGF-I or EGF stimulation. In the metastatic cancer tissues, KCC4 colocalizes with IGF-I or EGF, indicating a likely in vivo stimulation of KCC4 function by growth factors. Thus, blockade of KCC4 trafficking and surface expression may provide a potential target for the prevention of IGF-I- or EGF-dependent cancer spread.",
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Motor protein-dependent membrane trafficking of KCl cotransporter-4 is important for cancer cell invasion. / Chen, Yih Fung; Chou, Cheng-Yang; Wilkins, Robert J.; Ellory, J. Clive; Mount, David B.; Shen, Meng-Ru.

In: Cancer Research, Vol. 69, No. 22, 15.11.2009, p. 8585-8593.

Research output: Contribution to journalArticle

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AU - Chou, Cheng-Yang

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AU - Ellory, J. Clive

AU - Mount, David B.

AU - Shen, Meng-Ru

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