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Inhibition of CD44 expression in hepatocellular carcinoma cells enhances apoptosis, chemosensitivity, and reduces tumorigenesis and invasion

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Abstract

Purpose

CD44 is overexpressed in various tumors including hepatocellular carcinoma (HCC). The purpose of this study was to examine the effects of CD44 antisense oligonucleotide (ASO) alone or combination with doxorubicin on HCC cells in vitro.

Methods

Cytotoxicity was measured by use of a cell viability assay in HCC cell line SNU-449. Tumorigenesis and invasion were accessed by colony formation, growth in soft agar and ECMatrix invasion assay. Apoptosis and necrosis were evaluated by using double staining with Hoechst 33342 and propidium iodide. Protein expression and mRNA level were detected by Western blot and RT-PCR.

Results

We have designed novel CD44 ASO, which can effectively down-regulate CD44 expression in SNU-449. Colony formation, growth in soft agar and invasion were significantly impaired after CD44 ASO treatment in SNU-499. In company with CD44 down-regulated by CD44 ASO, MDR-1 and Bcl-2 expression were also greatly reduced. CD44 ASO also increased chemosensitivity to doxorubicin significantly, lowered IC50 by one order of magnitude. Apoptosis and necrosis were also induced by CD44 ASO alone or in combination treatment with doxorubicin.

Conclusions

Inhibition of CD44 expression by CD44 ASO significantly induced apoptosis, decreased tumorigenesis and invasion, and increased chemosensitivity. Thus, CD44 ASO is potentially a therapy that is worth investigating in the clinical setting.

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Acknowledgments

This study was supported by Singapore Cancer Syndicate Grant—TN0031, AN0038 and Terry Fox Run Cancer Research Grant 2004 (Chien-Shing Chen).

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Authors and Affiliations

  1. Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 119074, Singapore

    Zhigang Xie, Pei Feng Choong, Lai Fong Poon, Jianbiao Zhou, Jiaying Khng, Viraj Janakakumara Jasinghe, Senthilnathan Palaniyandi & Chien-Shing Chen

  2. Oncology Research Institute, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117456, Singapore

    Chien-Shing Chen

  3. Division of Hematology and Oncology, Loma Linda University, School of Medicine, Loma Linda, CA, 92354, USA

    Chien-Shing Chen

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  1. Zhigang Xie
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Correspondence to Chien-Shing Chen.

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Xie, Z., Choong, P.F., Poon, L.F. et al. Inhibition of CD44 expression in hepatocellular carcinoma cells enhances apoptosis, chemosensitivity, and reduces tumorigenesis and invasion. Cancer Chemother Pharmacol 62, 949–957 (2008). https://doi.org/10.1007/s00280-008-0684-z

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  • DOI: https://doi.org/10.1007/s00280-008-0684-z

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