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Targeting glucosylceramide synthase downregulates expression of the multidrug resistance gene MDR1 and sensitizes breast carcinoma cells to anticancer drugs

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Abstract

Drug resistance in breast cancer remains a major cause for the failure of chemotherapy. Glucosylceramide synthase (GCS) plays an important role in multidrug resistance (MDR) in breast cancer. P-glycoprotein (P-gp) also confers a cross-resistance of many unrelated drugs. In this study, we studied the MDR effect and potential mechanisms of breast cancer after constructing permanent breast cancer cell lines with GCS knockout by using recombinant vectors targeting GCS (pSUPER-GCSshRNAs). The GCSshRNA stably transfected cells were successfully established and significant lower levels of GCS mRNA and protein expression were confirmed. In in vitro experiments, the GCSshRNA stably transfected cells showed a significantly reduced level of MDR1 and P-gp expression and decreased drug efflux ability. Reduced level of GCS expression conveyed a significant reversal of drug resistance by MTT assay and increased caspase-3 activity. In in vivo experiments by using nude mice with xenograft tumors, a significant inhibition of tumor growth was observed after comparing with the control group. Furthermore, enhanced response of chemotherapy was acquired by reduced expression of GCS as well as MDR1 in vivo. In conclusion, GCSshRNA could efficiently suppress GCS and MDR1 expression in vitro and in vivo and these findings may be used as one of the methods to reverse MDR in breast cancer.

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Abbreviations

GCS:

Glucosylceramide synthase

MDR:

Multidrug resistance

P-gp:

P-glycoprotein

RNAi:

RNA interference

siRNA:

Small interfering RNA

shRNA:

Short hairpin RNA

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Acknowledgments

This work was supported in part by National Nature Science Foundation of China (No. 30300124); Nature Science Foundation of Shandong Province of China (No. Y2007C112, Y2007C15).

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, Shandong University, Jinan, 250012, People’s Republic of China

    Yanlin Sun, Tingguo Zhang, Peng Gao, Bin Meng, Xiao Wang, Jianping Zhang, Hao Wang, Xiaojuan Wu & Gengyin Zhou

  2. Department of Pathology, Shandong Tumor Hospital, Jinan, 250117, People’s Republic of China

    Yongsheng Gao

  3. Department of Pathology, University of Arizona College of Medicine, Tucson, AZ, USA

    Wenxin Zheng

Authors
  1. Yanlin Sun
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  2. Tingguo Zhang
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  3. Peng Gao
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  4. Bin Meng
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  5. Yongsheng Gao
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  6. Xiao Wang
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  7. Jianping Zhang
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  8. Hao Wang
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  9. Xiaojuan Wu
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  10. Wenxin Zheng
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  11. Gengyin Zhou
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Corresponding authors

Correspondence to Yanlin Sun or Gengyin Zhou.

Additional information

Dr. Yanlin Sun and Dr. Gengyin Zhou contributed equally to this study.

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Sun, Y., Zhang, T., Gao, P. et al. Targeting glucosylceramide synthase downregulates expression of the multidrug resistance gene MDR1 and sensitizes breast carcinoma cells to anticancer drugs. Breast Cancer Res Treat 121, 591–599 (2010). https://doi.org/10.1007/s10549-009-0513-z

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  • DOI: https://doi.org/10.1007/s10549-009-0513-z

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