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Small molecule inhibitor of c-Met (PHA665752) suppresses the growth of ovarian cancer cells and reverses cisplatin resistance

  • Original Article
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Tumor Biology

Abstract

c-Met as a tyrosine-kinase receptor plays a major role in tumorigenesis, invasion, and metastatic spread of human tumors, including ovarian cancer. Expressing high levels of c-Met proteins is often associated with resistance to chemotherapy and an adverse prognosis. In this study, we have determined the effect of PHA665752, a small molecule inhibitor of c-Met proteins, with and without cisplatin and the role of c-Met in several ovarian cancer cell lines having high c-Met expression. The methyl thiazolyl tetrazolium (MTT) assay was used to detect cell proliferation, and apoptosis was evaluated by flow cytometry. Western blotting was carried out to determine protein expression levels. Gene silencing was used to detect the influence of c-Met gene silence on the resistance to cisplatin. Compared to more sensitive ovarian cancer cell lines SKOV3 and 3AO, we found that the expression of c-Met was significantly increased in SKOV3DDP, OVCAR3, and OV-90 ovarian cancer cell lines, which were resistant to cisplatin. Our data indicated that cisplatin sustained activated phosphor-Met in SKOV3DDP, OVCAR3, and OV-90 cell lines. We also observed a significant transient activation of c-Met phosphorylation in SKOV3 and 3AO cells. Treatment with PHA665752 inhibited c-Met expression inhibited cell growth, induced apoptosis, and enhanced cisplatin-induced proliferation inhibition and apoptosis in c-Met over-expressed cell lines. In addition, blocking c-Met expression with small interfering RNA (siRNA) overcame the resistance of cancer cells to cisplatin. Thus, blocking c-Met expression presents a promising therapeutic approach for ovarian cancer.

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Authors and Affiliations

  1. Laboratory of Sono- and Photo-theranostic Technologies, Harbin Institute of Technology, Harbin, 150080, China

    Enze Li, Zheng Hu, Qi Zhou, Bin Yang & Wenwu Cao

  2. Department of Stomatology, The Fourth Affiliated Hospital, Harbin Medical University, Harbin, 150000, China

    Yi Sun

  3. Condensed Matter Science and Technology Institute, Harbin Institute of Technology, Harbin, 150080, China

    Zhiguo Zhang

  4. Department of Mathematics and Materials Research Institute, Pennsylvania State University, 164 Materials Research Lab, University Park, PA, 16802, USA

    Wenwu Cao

Authors
  1. Enze Li
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  2. Zheng Hu
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  3. Yi Sun
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  4. Qi Zhou
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  5. Bin Yang
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  6. Zhiguo Zhang
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  7. Wenwu Cao
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Corresponding authors

Correspondence to Zhiguo Zhang or Wenwu Cao.

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Funding support

This work was supported by grants from the National Key Technology R&D Program of China (Grant No. 2013BAI03B06), the National Natural Science Foundation of China (Grant No. 81272503), the Fundamental Research Funds for the Central Universities (Grant No. HIT.NSRIF.2014066), and the China Postdoctoral Science Foundation (Grant No. 2014 M561341).

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Supplemental Figure 7

Effect of PHA665752 on apoptosis in 3AO and SKOV3 cells. 3AO and SKOV3 cells were exposed to PHA665752 (400nmol/L) for 24-48 h and then the percentage of apoptotic cells was analyzed by flow cytometry after staining with PI. Sub-G1 phase represents early apoptosis cells. Data are means ± SD of three independent experiments. (GIF 232 kb)

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Li, E., Hu, Z., Sun, Y. et al. Small molecule inhibitor of c-Met (PHA665752) suppresses the growth of ovarian cancer cells and reverses cisplatin resistance. Tumor Biol. 37, 7843–7852 (2016). https://doi.org/10.1007/s13277-015-4318-x

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  • DOI: https://doi.org/10.1007/s13277-015-4318-x

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