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Chemoresistance to Temozolomide in Human Glioma Cell Line U251 is Associated with Increased Activity of O 6-methylguanine-DNA Methyltransferase and Can be Overcome by Metronomic Temozolomide Regimen

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Abstract

Temozolomide (TMZ) is a novel cytotoxic alkylating agent for chemotherapy of malignant gliomas. However, intrinsic or acquired resistance to TMZ often defines poor efficacy of chemotherapy in malignant gliomas. A growing number of studies indicate that expression of O 6-methylguanine-DNA methyltransferase (MGMT) is one of the principal mechanisms responsible for this chemoresistance. In the present study, we evaluated the relationship between expression of MGMT and resistance to TMZ. We generated a TMZ-resistant cell line, U251/TR, by stepwise (8 months) exposure of parental U251 cells to TMZ. The resistance to TMZ was quantified using SRB assay. MGMT expression was evaluated at mRNA (RT-PCR) and protein (Western blot) levels. U251/TR cells showed increased (~ sevenfold) resistance to TMZ. The MGMT expression (both mRNA and protein) was significantly (P < 0.01) increased in U251/TR cells compared with parental U251 cells. Further, MGMT expression fluctuated during exposure of U251/TR cells to TMZ. The resistance of U251/TR cells to TMZ could be overcome by application of elevated doses of TMZ when MGMT expression was at the lowest level. In conclusion, our results demonstrate that the primary mechanism responsible for resistance of U251/TR cells to TMZ is associated with increased expression of MGMT. Resistance of malignant gliomas to TMZ can be overcome by synchronizing metronomic TMZ regimen with MGMT expression.

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Acknowledgments

The authors gratefully acknowledge the financial support by the National Natural Science Foundation of China (grants no. 30571904 and 30772228).

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Authors and Affiliations

  1. Department of Neurosurgery, General Hospital, Tianjin Medical University, 154 An-Shan Road, Heping District, Tianjin, 300052, People’s Republic of China

    Qiang Pan, Xue-jun Yang, Hua-min Wang, Xue-tao Dong, Wei Wang & Yu LI

  2. Neuro-Oncology Laboratory, Tianjin Neurological Institute, Tianjin, People’s Republic of China

    Qiang Pan, Hua-min Wang, Xue-tao Dong, Wei Wang & Yu LI

  3. Institute of Human Anatomy and Embryology, Bin Zhou Medical College, Bin Zhou, People’s Republic of China

    Jing-min LI

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  1. Qiang Pan
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  2. Xue-jun Yang
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  4. Xue-tao Dong
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  5. Wei Wang
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Correspondence to Xue-jun Yang.

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Qiang Pan and Xue-jun Yang are contributed equally.

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Pan, Q., Yang, Xj., Wang, Hm. et al. Chemoresistance to Temozolomide in Human Glioma Cell Line U251 is Associated with Increased Activity of O 6-methylguanine-DNA Methyltransferase and Can be Overcome by Metronomic Temozolomide Regimen. Cell Biochem Biophys 62, 185–191 (2012). https://doi.org/10.1007/s12013-011-9280-7

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  • DOI: https://doi.org/10.1007/s12013-011-9280-7

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