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Chloride intracellular channel 4 involves in the reduced invasiveness of cancer cells treated by photodynamic therapy

Authors

  • Pei-Chi Chiang MS,

    1. Department of Biochemical Science and Technology, National Taiwan University, Taipei 10617, Taiwan
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  • Ruey-Hwang Chou PhD,

    1. Graduate Institute of Cancer Biology and Center for Molecular Medicine, China Medical University, Taichung 404, Taiwan
    2. Department of Biotechnology, Asia University, Taichung 413, Taiwan
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  • Hsiung-Fei Chien MD, PhD,

    1. Department of Surgery, National Taiwan University College of Medicine, Taipei 100, Taiwan
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  • Tsuimin Tsai PhD,

    1. Graduate Institute of Biomedical Materials and Tissue Engineering, College of Oral Medicine, Taipei Medical University, Taipei 11031, Taiwan
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  • Chin-Tin Chen PhD

    Corresponding author
    1. Department of Biochemical Science and Technology, National Taiwan University, Taipei 10617, Taiwan
    • Department of Biochemical Science and Technology, College of Life Science, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei 10617, Taiwan.
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  • Conflict of Interest Disclosures: All authors have completed and submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest and none were reported.

  • Ruey-Hwang Chou and Hsiung-Fei Chien contributed equally to this work.

Abstract

Background and Objectives

The mechanisms of photodynamic therapy (PDT) have been studied on the cellular and tissue levels. However, the cellular behaviors of cancer cells survived from PDT are still not clear. Previously, we have found that PDT-derived variants A375/3A5 and A375/6A5 have reduced invasion ability. This study attempted to further elucidate the possible molecules associated with the altered invasiveness in the PDT-derived variants and cancer cells treated with PDT.

Study Design/Materials and Methods

Scratch wound healing assay and invasion assay were performed to evaluate the migration and invasion ability of human A375 melanoma and MDA-MB-231 breast adenocarcinoma cells. Single colony selection and microarray analysis were performed to examine the differentially expressed transcripts in parental A375 and PDT-derived variants. RT-PCR and Western blots analysis were performed to examine the expression levels of matrix metalloproteinase 9 (MMP9) and chloride intracellular channel 4 (CLIC4). The MMP9 activity was examined by Zymography assay. CLIC4 expressing construct was used to examine the influence on MMP9 expression and invasion ability of cancer cells treated with PDT.

Results

Correlated with the reduced invasiveness, we found that A375/3A5 and A375/6A5 cells have decreased production of MMP9. Microarray analysis and RT-PCR showed CLIC4 was down-regulated in the PDT-derived variants. Furthermore, down-regulation of CLIC4 and MMP9 was found in cancer cells treated with PDT. Transfection of surviving cancer cells with a plasmid vector encoding CLIC4 increased MMP9 expression and cell invasion. Furthermore, overexpression of CLIC4 in A375 and MDA-MB-231 cancer cells constrains PDT-induced suppression of invasiveness.

Conclusion

Our results showed that the reduced expression of CLIC4 could further down-regulate MMP9 and result in the suppression of invasion in cancer cells treated with PDT. These results provide an insight into a new mechanism by which PDT affects the metastatic potential of cancer cells through down-regulation of MMP9 by CLIC4. Lasers Surg. Med. 45: 38–47, 2013. © 2013 Wiley Periodicals, Inc.

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