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Status of p53 phosphorylation and function in sensitive and resistant human cancer models exposed to platinum-based DNA damaging agents

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Abstract

Purpose

Resistance to chemotherapeutic drugs is a hallmark of many human cancers, which can occur independent of p53 gene status; however, the presence of wild-type p53 in chemorefractory tumors confers greater resistance to cisplatin, but such tumors do not display complete cross-resistance to the platinum analog (1R,2R-diaminocyclohexane)(trans-diacetato)(dichloro)platinumIV (DACH-Ac-Pt). In this article we examine DNA damage-induced phosphorylation of p53 and downstream p53-dependent transactivation events in cisplatin-sensitive and cisplatin-resistant human cancer cell lines possessing wild-type p53.

Methods

Western-blot analysis was utilized to study the effect of cisplatin and the analog on p53 phosphorylation and p53-dependent target genes.

Results

In response to CDDP and DACH-Ac-Pt, both CDDP-sensitive and CDDP-resistant models demonstrated time- and dose-dependent inductions of total p53 protein and an increase in Ser-15 phosphorylation, which was more pronounced with CDDP. Although phosphorylation of p53 at Ser-392 was also observed in CDDP-treated sensitive and resistant cells, it was weak or absent in response to DACH-Ac-Pt. Lack of Ser-392 phosphorylation by DACH-Ac-Pt, however, did not affect the induction of p21WAF1/CIP1 or Mdm2. Similarly, inductions of p21WAF1/CIP1 and Mdm2 were observed in sensitive cells exposed to cisplatin. In marked contrast, cisplatin-mediated induction of p21WAF1/CIP1 was minimal or absent in resistant cells, but that of Mdm2 was unaffected. Wortmannin, a PI3-kinase (PI3-K) inhibitor, caused a dose-dependent inhibition of total p53 accumulation, Ser-15 phosphorylation and p21WAF1/CIP1 transactivation in response to both CDDP and DACH-Ac-Pt, indicating that members of the PI3-K family are involved in phosphorylation of p53 and that transactivation of p21WAF1/CIP1 is p53 dependent.

Conclusion

These studies demonstrate that cisplatin and DACH-Ac-Pt differentially phosphorylate p53 through independent DNA damage-induced pathways, and that the kinase-mediated phosphorylation of p53 at Ser-15 or Ser-392 is unaltered in resistance. Moreover, the phosphorylation status of Ser-392 on its own does not appear to correlate with p21WAF1/CIP1 or Mdm2 induction in these studies; however, a lack of increase in p21WAF1/CIP1 by cisplatin, but not DACH-Ac-Pt, provides a correlation with resistance and its circumvention, and implicates the role for cyclin-dependent kinase inhibitor in the differential cytotoxic effects of the two platinum agents against resistant cells.

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Acknowledgement. The authors thank P. Hennessey for his expert technical assistance, as well as K. Biescar and J. Neicheril for preparation of the manuscript. This work was supported by NCI RO1 CA77332 and CA82361, and U.S. Army DAMD 17-99-1-9269 grants to Z.H.S.

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  1. Junichi Nakamura

    Present address: Department of Surgery, Saitama Red Cross Hospital, 8-3-33 Kamiochiai, Chuuou, 338-8553, Saitama City, Japan

  2. Masayuki Watanabe

    Present address: Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-Ku, 812-8582, Fukuoka, Japan

Authors and Affiliations

  1. Department of Experimental Therapeutics, The University of Texas M.D. Anderson Cancer Center, Box 104, Houston, TX, 77030, USA

    Kalpana Mujoo, Masayuki Watanabe, Junichi Nakamura, Abdul R. Khokhar & Zahid H. Siddik

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  1. Kalpana Mujoo
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  2. Masayuki Watanabe
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Correspondence to Zahid H. Siddik.

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Mujoo, K., Watanabe, M., Nakamura, J. et al. Status of p53 phosphorylation and function in sensitive and resistant human cancer models exposed to platinum-based DNA damaging agents. J Cancer Res Clin Oncol 129, 709–718 (2003). https://doi.org/10.1007/s00432-003-0480-4

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  • DOI: https://doi.org/10.1007/s00432-003-0480-4

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