Modulation of excision repair cross complementation group 1 (ercc-1) mrna expression by pharmacological agents in human ovarian carcinoma cells

doi:10.1016/S0006-2952(98)00291-3

Biochemical Pharmacology

Volume 57, Issue 4, 15 February 1999, Pages 347-353

https://doi.org/10.1016/S0006-2952(98)00291-3 Get rights and content

Abstract

Excision repair cross complementation group 1 (ERCC-1) is a DNA repair gene that is essential for life, and it appears to be a marker gene for nucleotide excision repair activity. Overexpression of ERCC-1 during cisplatin-based chemotherapy is associated with clinical and cellular drug resistance. We therefore began to assess the influence of various pharmacological agents on the induction of ERCC-1 mRNA in A2780/CP70 human ovarian carcinoma cells. Cisplatin exposure in culture resulted in a 4- to 6-fold induction for the steady-state level of ERCC-1 mRNA in A2780/CP70 cells. ERCC-1 mRNA induction was concentration and time dependent. Cyclosporin A and herbimycin A, which suppress c-fos and c-jun gene expressions, respectively, blocked the cisplatin-induced increase in ERCC-1 mRNA. This effect of cyclosporin A or herbimycin A on the down-regulation of ERCC-1 correlates with enhanced cytotoxicity of cisplatin in this system. The products of c-fos and c-jun are components of the transcription factor AP-1 (activator protein 1). 12-O-Tetradecanoylphorbol 13-acetate (TPA), a known AP-1 agonist, induced ERCC-1 mRNA to the same extent as cisplatin, but did not synergize with cisplatin in this regard. The TPA effect was biphasic, with an initial increase during the first 1–6 hr, followed by decreasing mRNA levels at 24–72 hr. These data suggest that the effects of these pharmacological agents on ERCC-1 gene expression may be mediated through the modulation of AP-1 activities.

Section snippets

Cell line and cell culture conditions

The human ovarian cancer cell line A2780/CP70 has been described previously [24] and was used in all the experiments. Cells were cultured in monolayer using RPMI 1640 medium supplemented with 10% (v/v) fetal bovine serum, 2 mM l-glutamine, 0.2 U/mL of human insulin, 50 U/mL of penicillin, and 50 μg/mL of streptomycin (Gibco BRL). Cells were grown logarithmically at 37° in a humidified atmosphere consisting of 5% CO₂:95% air. Cells were tested routinely for mycoplasmal infection, using a

Up-regulation of ERCC-1 gene expression by cisplatin in human ovarian carcinoma A2780/CP70 cells

Previous studies from our laboratory have shown that cisplatin-resistant human ovarian cancer tissues show an increased expression of ERCC-1 mRNA 21, 23. In the present study, we first determined whether cisplatin can directly stimulate ERCC-1 gene expression in a human ovarian cancer cell line, A2780/CP70. The A2780/CP70 cell line was treated with 40 μM cisplatin for 1 hr, and the expression of ERCC-1 mRNA was measured at various time points following drug exposure. Cisplatin caused time- and

Discussion

NER is the principal DNA repair pathway by which cisplatin damage, as well as other types of bulky DNA lesions, is removed from cellular DNA 13, 14, 15, 16, 17. ERCC-1 is one of the critical genes within NER 32, 33, 34 and may be responsible for the incision 5′ to the site of DNA damage [35]. ERCC-1 gene expression is necessary for the repair of cisplatin-induced DNA damage [13], and enhanced expression is associated with clinical resistance to cisplatin 21, 23. Thus, understanding the

Acknowledgements

We thank Dr. Christopher T. Bever for advice. Dr. Qingdi Li was supported by a postdoctoral intramural research training award (IRTA) from the National Institutes of Health.

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ERCC1 is often discussed as a predictive marker of cisplatin response [10]. Regarding expression of ERCC1 mRNA upon CDDP treatment, increased ERCC1 mRNA levels have been reported in ovarian carcinoma cells [64,65] while decreased ERCC1 mRNA was reported in lung cancer cells [66] suggesting a cell-type specific response to the platinum drug. CDKN1A-encoded p21 is an important regulator of cell cycle progression leading to G1, G2 or S-phase arrest [43–46] while the protein phosphatase PPM1D and the serine-threonine protein kinase PLK3 affect the G1/S and G2/M blockade [39–42].
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L’excision repair cross-complementation group 1 (ERCC1) joue un rôle critique dans la réparation des lésions induites sur l’ADN par le cisplatine. Il a été démontré que les patients avec un cancer bronchique non à petites cellules (CBNPC) opéré et dont les tumeurs sont ERCC1-négatives ont un bénéfice plus important de la chimiothérapie adjuvante par rapport aux patients ERCC1-positifs. En revanche, les patients dont les tumeurs sont ERCC1-positives ont un meilleur pronostic. L’étude d’ERCC1 peut être faite soit par la mesure de son expression protéique par immunohistochimie (anticorps commercialisés) soit par quantification de son expression transcriptionnelle (mesure de l’ARNm par RT-PCR). ERCC1 apparaît ainsi comme un biomarqueur prometteur pour prédire le bénéfice de la chimiothérapie à base de cisplatine et sa modulation pourrait permettre de sensibiliser les cellules tumorales. Son utilisation en pratique clinique quotidienne nécessite encore une étape de validation prospective dont des études sont en cours soit chez les patients opérés d’un CBNPC, soit au stade métastatique.
ERCC1 plays a critical role in the repair of lesions in DNA induced by cisplatin. It has been demonstrated that, in patients with resected NSCLC, those with ERCC1-negative tumours had greater benefit from adjuvant chemotherapy than patients with ERCC1-positive tumours. On the other hand, patients with ERCC1-positive tumours have a better prognosis. ERCC1 can be evaluated either by the level of protein expression on immunohistochemical analysis or by quantification of its mRNA expression. At present ERCC1 appears to be a promising biomarker for predicting the benefit of cisplatin based chemotherapy and its modulation could allow sensitisation of tumour cells to this drug. Its use in daily clinical practice requires a further stage of prospective validation studies which are already in progress in patients both with completely resected NSCL or metastatic disease.
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Cyclosporin A has been shown to suppress expression of the c-fos gene which may in turn suppress the expression of ERCC1 mRNA induced by cisplatin.89 Herbimycin A, a protein kinase A inhibitor has been demonstrated to suppress cisplatin-induced c-jun mRNA expression.89 While the data presented above show that dysfunctional DNA repair in tumour tissue may have an impact on anti-cancer activity of DNA damaging agents, reduced DNA repair capacity in normal tissue may constitute a risk factor for cancer development.90,91
Cisplatin, carboplatin and oxaliplatin are some of the most widely used anti-cancer agents in solid tumours. The cytotoxicity of platinating agents is directly related to their ability to cause DNA intra-strand crosslinks that trigger a series of intracellular events that ultimately result in cell death. DNA intra-strand crosslinks are processed and repaired by the nucleotide excision repair pathway. It is now clear that nucleotide excision repair (NER) capacity may have a major impact on the emergence of resistance, normal tissue tolerance and patient outcomes. ERCC1 is a key player in NER. In this review, we provide an overview of mammalian NER and then focus on biochemical, structural and pre-clinical aspects of ERCC1. We then present current clinical evidence implicating ERCC1 as a predictive and prognostic marker in cancer. Early evidence also suggests that ERCC1 or the pathways involved in the regulation of ERCC1 expression may be attractive anti-cancer targets. Such agents are expected to potentiate the cytotoxicity of platinating agents and could have a major impact on cancer therapy.
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Molecular and Cellular PharmacologiesModulation of excision repair cross complementation group 1 (ercc-1) mrna expression by pharmacological agents in human ovarian carcinoma cells

Abstract

Section snippets

Cell line and cell culture conditions

Up-regulation of ERCC-1 gene expression by cisplatin in human ovarian carcinoma A2780/CP70 cells

Discussion

Acknowledgements

J Biol Chem

J Biol Chem

Toxicol In Vitro

Anal Biochem

J Biol Chem

J Biol Chem

Cell

Cell

Cell

Cancer Treat Rev

The chemotherapy of ovarian cancer

PPO Update

Interaction of platinum antitumor compounds with DNA

Nucleic Acids Mol Biol

Acquired cisplatin resistance in human ovarian cancer cells is associated with enhanced DNA repair of cisplatin–DNA lesions and reduced drug accumulation

J Clin Invest

Cisplatin

Cancer Chemother Biol Response Modif

Insights into mechanisms of cisplatin resistance and potential for its clinical reversal

Pharmacotherapy

Positive correlation between cellular glutathione and acquired cisplatin resistance in human ovarian cancer cells

Cell Biol Toxicol

High resistance to cisplatin in human ovarian cancer cell lines is associated with marked increase of glutathione synthesis

Proc Natl Acad Sci USA

Cross-resistance to diverse drugs is associated with primary cisplatin resistance in ovarian cancer cell lines

Cancer Res

Increased DNA repair as a mechanism of acquired resistance to cis-diamminedichloroplatinum (II) in human ovarian cancer cell lines

Cancer Res

Molecular and Cellular Pharmacologies
Modulation of excision repair cross complementation group 1 (ercc-1) mrna expression by pharmacological agents in human ovarian carcinoma cells