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HuR-targeted nanotherapy in combination with AMD3100 suppresses CXCR4 expression, cell growth, migration and invasion in lung cancer

Cancer Gene Therapy volume 22, pages 581–590 (2015)Cite this article

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Abstract

The CXCR4 chemokine receptor has an important role in cancer cell metastasis. The CXCR4 antagonist, AMD3100, has limited efficacy in controlling metastasis. HuR, an RNA-binding protein, regulates CXCR4 in cancer cells. We therefore investigated whether targeting HuR using a siRNA-based nanoparticle plus AMD3100 would suppress CXCR4 and inhibit lung cancer metastasis. We treated human H1299 lung cancer cells with HuR-specific siRNA contained in a folate-targeted lipid nanoparticle (HuR-FNP) plus AMD3100, and compared this with AMD3100 alone, HuR-FNP alone and no treatment. HuR-FNP plus AMD3100 treatment produced a G1 phase cell cycle arrest and reduced cell viability above and beyond the effects of AMD3100 alone. HuR and CXCR4 mRNA and protein expression levels were markedly reduced in all treatment groups. Phosphorylated (p) AKTS473 protein was also reduced. P27 protein expression increased with HuR-FNP and combination treatment. Promoter-based reporter studies showed that the combination inhibited CXCR4 promoter activity more than did either treatment alone. Cell migration and invasion was significantly reduced with all treatments; the combination provided the most inhibition. Reduced matrix metalloprotease (MMP)-2 and -9 expression was associated with reduced invasion in all treatment groups. Thus, we found that combined HuR and CXCR4 targeting effectively controlled lung cancer metastasis.

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Acknowledgements

The study was supported in part by a grant received from the National Institutes of Health (NIH) R01 CA167516, an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences (P20 GM103639) of the National Institutes of Health, and by funds received from the Jim and Christy Everest Endowed Chair in Cancer Developmental Therapeutics, the University of Oklahoma Health Sciences Center. We thank the Stephenson Cancer Center at the University of Oklahoma Health Sciences Center, Oklahoma City, OK, for the use of the cancer Functional Genomics Core and Biostatistics Core, which provided molecular analysis and statistical services. We thank Ms. Kathy Kyler at the office of Vice President of Research, OUHSC for editorial assistance. Rajagopal Ramesh is an Oklahoma TSET Research Scholar and holds the Jim and Christy Everest Endowed Chair in Cancer Developmental Therapeutics.

Author contributions

RM and JP conducted the experiments. RM synthesized and characterized the nanoparticle (HuR-FNP and C-FNP). RM, JP, AC, YDZ, AM and RR conceived and designed the experiments. RM and RR wrote the paper. AC and YDZ performed statistical analysis. Together, RM, JP, AC, YDZ, AM and RR did data collection, analysis and interpretation. RM, JP, AC, YDZ, AM and RR critically reviewed, revised for intellectual content, and provided suggestions. RR supervised the project.

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Authors and Affiliations

  1. Department of Pathology, Stanton L. Young Biomedical Research Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    R Muralidharan, J Panneerselvam & R Ramesh

  2. Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    R Muralidharan, J Panneerselvam, Y D Zhao, A Munshi & R Ramesh

  3. Department of Biostatistics and Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    A Chen & Y D Zhao

  4. Department of Radiation Oncology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    A Munshi

  5. Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    R Ramesh

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Correspondence to R Ramesh.

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Muralidharan, R., Panneerselvam, J., Chen, A. et al. HuR-targeted nanotherapy in combination with AMD3100 suppresses CXCR4 expression, cell growth, migration and invasion in lung cancer. Cancer Gene Ther 22, 581–590 (2015). https://doi.org/10.1038/cgt.2015.55

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