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miR-200bc/429 cluster modulates multidrug resistance of human cancer cell lines by targeting BCL2 and XIAP

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Abstract

Purpose

MicroRNAs (miRNAs) are short non-coding RNA molecules, which post-transcriptionally regulate genes expression and play crucial roles in diverse biological processes. Recent studies have shown that dysregulation of miRNAs might modulate the resistance of cancer cells to anti-cancer drugs, yet the modulation mechanism is not fully understood. We aimed to investigate the possible role of miRNAs in the development of multidrug resistance (MDR) in human gastric and lung cancer cell lines.

Methods

miRNA Quantitative real-time PCR was used to detect the different miRNA expression levels between drug resistant and parental cancer cells. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test the drug-resistant phenotype changes in cancer cells via over or downregulation of miRNAs. Dual-luciferase activity assay was used to verify the target genes of miRNAs. Western blot analysis and apoptosis assay were used to elucidate the mechanism of miRNAs on modulating drug resistance in cancer cells.

Results

miR-200bc/429 cluster was downregulated, while BCL2 and XIAP were upregulated in both MDR SGC7901/VCR (vincristine) and A549/CDDP (cisplatin) cells, compared with the parental SGC7901 and A549 cells, respectively. Overexpression of miR-200bc/429 cluster sensitized SGC7901/VCR and A549/CDDP cells to anti-cancer drugs, respectively. Both BCL2 and XIAP 3′-UTR reporters constructed in MDR cells suggested that BCL2 and XIAP were the common target genes of the miR-200bc/429 cluster. Enforced miR-200bc/429 cluster expression reduced BCL2 and XIAP protein level and sensitized both MDR cells to VCR-induced and CDDP-induced apoptosis, respectively.

Conclusions

Our findings first suggest that miR-200bc/429 cluster could play a role in the development of MDR in both gastric and lung cancer cell lines, at least in part by modulation of apoptosis via targeting BCL2 and XIAP.

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Abbreviations

miRNAs:

MicroRNAs

MDR:

Multidrug resistance

VCR:

Vincristine

CDDP:

Cisplatin

ADR:

Adriamycin

5-Fu:

5-Fluorouracil

VP-16:

Etoposide

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Acknowledgments

The authors are grateful to the fund supported by the National Natural Science Foundation of China (Grant number: 30840095) and the Cancer Center of Nanjing Medical University (Grant number: 08ZLKF02).

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Authors and Affiliations

  1. Department of Oncology, First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China

    Wei Zhu, Hui Zhi, Tongshan Wang, Jian Wang, Yongqian Shu & Ping Liu

  2. Cancer Center of Nanjing Medical University, 140 Hanzhong Road, Nanjing, 210029, China

    Binghua Jiang, Yongqian Shu & Ping Liu

  3. Department of Haematology, First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China

    DanXia Zhu

  4. Department of Clinical Laboratory, First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China

    Huaguo Xu

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  1. Wei Zhu
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  9. Ping Liu
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Correspondence to Ping Liu.

Additional information

Wei Zhu and Huaguo Xu contributed equally.

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Zhu, W., Xu, H., Zhu, D. et al. miR-200bc/429 cluster modulates multidrug resistance of human cancer cell lines by targeting BCL2 and XIAP. Cancer Chemother Pharmacol 69, 723–731 (2012). https://doi.org/10.1007/s00280-011-1752-3

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  • DOI: https://doi.org/10.1007/s00280-011-1752-3

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