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miR-181a sensitizes resistant leukaemia HL-60/Ara-C cells to Ara-C by inducing apoptosis

Abstract

Background

Ara-C is one of the most commonly used drugs in the treatment of AML. However, the development of drug resistance always prevented its further use. It has been shown that miR-181a is associated with the clinical outcome of AML patients. Here, we investigated the possible role of miR-181a in AML Ara-C resistance.

Methods

miR-181a expression was measured by real-time PCR. Cell viability was detected by MTT assay. Protein expressions were measured by western blotting. Caspase activity was examined by florescence assay.

Results

We found that miR-181a expression was downregulated in the Ara-C-resistant cell line HL-60/Ara-C compared with its parental cell line HL-60. Overexpression of miR-181a in HL-60/Ara-C cells sensitized the cells to Ara-C treatment. Furthermore, Bcl-2 was confirmed as a direct miR-181a target by immunoblot analysis and reporter gene assays. Knockdown of Bcl-2 mimicked the effect of enforced miR-181a expression by reducing cell viability. In addition, the apoptosis pathway was activated by cytochrome C release and caspase 9/caspase 3 activation after miR-181a overexpression.

Conclusions

This study for the first time demonstrated that downregulation of miR-181a and upregulation of Bcl-2 in leukaemia cells confer resistance to Ara-C-based therapy. These results suggest that restoration of miR-181a expression might provide a promising therapeutic in drug resistance of leukaemia.

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Acknowledgments

We thank L. Su for reviewing the manuscript. This study was supported by grants from Shanghai Jiaotong University Affiliated Shanghai First People’s Hospital (061138).

Conflict of interest

We declare that we have no conflict of interest.

Author information

Correspondence to Haitao Bai.

Additional information

Haitao Bai, Zhongwei Cao and Chong Deng contributed equally to this work.

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Bai, H., Cao, Z., Deng, C. et al. miR-181a sensitizes resistant leukaemia HL-60/Ara-C cells to Ara-C by inducing apoptosis. J Cancer Res Clin Oncol 138, 595–602 (2012). https://doi.org/10.1007/s00432-011-1137-3

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Keywords

  • Chemoresistance
  • miR-181a
  • Ara-C
  • Acute myeloid leukaemia
  • Bcl-2