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Article

Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells

  • Authors:
    • Xiao-You Li
    • Jian-Zhong Wu
    • Hai-Xia Cao
    • Rong Ma
    • Jian-Qiu Wu
    • Yue-Jiao Zhong
    • Ji-Feng Feng
  • View Affiliations / Copyright

    Affiliations: Department of Chemotherapy, Jiangsu Cancer Hospital and Research Institute, Nanjing, Jiangsu 210009, P.R. China
  • Pages: 1975-1982
    |
    Published online on: February 21, 2013
       https://doi.org/10.3892/or.2013.2298
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Abstract

The sensitivity of lung cancer to epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) has been found to be associated with mutations in the tyrosine kinase domain of EGFR. However, not all mutations are sensitive to gefitinib. While CpG island methylation in the promoter region of the EGFR gene and transcriptional silencing are common in solid tumors, the role of the EGFR gene promoter methylation in affecting resistance to TKIs in non-small cell lung cancer (NSCLC) remains unknown. In this study, we examined the correlation between EGFR gene promoter methylation and the therapeutic effect of gefitinib in NSCLC cells. Three NSCLC cell lines with different EGFR mutation statuses and levels of sensitivity to EGFR-TKIs were used in this study: H1650 (del E746-A750), H1299 (wild-type EGFR) and PC-9 (del E746-A750). Cells were treated with gefitinib or 5-aza-2'-deoxy cytidine (5-aza-CdR), a methylation inhibitor, alone or in combination. Subsequently, the methylation status of the EGFR gene promoter was examined by methylation-specific PCR (MSP). Cell survival and apoptosis assays were performed using the Cell Counting Kit-8 (CCK-8) and flow cytometry. In addition, western blot analysis and quantitative real-time PCR were used to examine the expression levels of EGFR protein and mRNA. Our study showed that the promoter region of the EGFR gene in PC-9 cells was unmethylated, and that the cells were sensitive to gefitinib. By contrast, the promoter region of the EGFR gene in the H1650 and H1299 cells was methylated, and the cells were resistant to gefitinib. Of note, the combination treatment with 5-aza-CdR and gefitinib further enhanced the growth inhibitory effects and led to the induction of apoptosis, while a significant reduction in the expression of EGFR protein and mRNA was observed in the H1650 and H1299 cells. These results suggest that blockade of DNA methylation may enhance the antitumor effects of EGFR-TKIs and gefitinib in NSCLC cells. Thus, EGFR gene promoter methylation may be a potential mechanism for acquired resistance to gefitinib.
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Copy and paste a formatted citation
Spandidos Publications style
Li X, Wu J, Cao H, Ma R, Wu J, Zhong Y and Feng J: Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells. Oncol Rep 29: 1975-1982, 2013.
APA
Li, X., Wu, J., Cao, H., Ma, R., Wu, J., Zhong, Y., & Feng, J. (2013). Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells. Oncology Reports, 29, 1975-1982. https://doi.org/10.3892/or.2013.2298
MLA
Li, X., Wu, J., Cao, H., Ma, R., Wu, J., Zhong, Y., Feng, J."Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells". Oncology Reports 29.5 (2013): 1975-1982.
Chicago
Li, X., Wu, J., Cao, H., Ma, R., Wu, J., Zhong, Y., Feng, J."Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells". Oncology Reports 29, no. 5 (2013): 1975-1982. https://doi.org/10.3892/or.2013.2298
Copy and paste a formatted citation
x
Spandidos Publications style
Li X, Wu J, Cao H, Ma R, Wu J, Zhong Y and Feng J: Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells. Oncol Rep 29: 1975-1982, 2013.
APA
Li, X., Wu, J., Cao, H., Ma, R., Wu, J., Zhong, Y., & Feng, J. (2013). Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells. Oncology Reports, 29, 1975-1982. https://doi.org/10.3892/or.2013.2298
MLA
Li, X., Wu, J., Cao, H., Ma, R., Wu, J., Zhong, Y., Feng, J."Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells". Oncology Reports 29.5 (2013): 1975-1982.
Chicago
Li, X., Wu, J., Cao, H., Ma, R., Wu, J., Zhong, Y., Feng, J."Blockade of DNA methylation enhances the therapeutic effect of gefitinib in non-small cell lung cancer cells". Oncology Reports 29, no. 5 (2013): 1975-1982. https://doi.org/10.3892/or.2013.2298
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