Role of membrane folate-binding protein in the cytotoxicity of 5,10-dideazatetrahydrofolic acid in human ovarian carcinoma cell lines in vitro

S. Sen, E. Erba, M. D'Incalci, F. Bottero, S. Canevari, A. Tomassetti

Research output: Contribution to journalArticle

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Abstract

Lometrexol (5,10-dideazatetrahydrofolic acid; DDATHF), is a specific inhibitor of glycinamideribonucleosyl (GAR) transformylase with anti-tumour activity in murine and human carcinomas. The cytotoxicity activity of DDATHF was evaluated in vitro in NIH/3T3 cells transfected with human α-folate-binding protein (FBP) complementary DNA to examine the role of the receptor. In FBP-transfecled NIH/3T3 (FBP-tNIH/3T3) cells, which internalised about three times more 5-methyltetrahydrofolic acid than the mock-transfected cells, the cytotoxic potential of DDATHF showed a clear increase. Subsequently, we analysed four ovarian carcinoma cell lines (OVCAR3, IGROV1, SKOV3 and SW626) expressing different amounts of FBP. Cells were conditioned to grow in medium depleted of folic acid then tested by MOv18 and folic acid binding. Only SKOV3 and SW626 cells grown in folic acid-depleted medium showed increased FBP expression, about 3-and 8-fold respectively. The cytotoxic potential of DDATHF was evaluated by a standard clonogenic assay. In a medium containing 2.27 μM folic acid the DDATHF IC50 values were 50 nM on OVCAR3, 500 nM on SW626 and 1000 nM on IGROV1. In folic acid-free medium IC50 values were 2 nM on OVCAR3 and SW626 and 40 nM on IGROV1. Only on SKOV3 cells was DDATHF cytotoxicity the same regardless of the amount of folic acid in the medium (IC50 8 nM). Thus, DDATHF did not inhibit the growth of IGROV1 cells depleted of folic acid after stripping FBP with phosphatidylinositol-phospholipase C, even at a dose toxic for cells constitutively expressing FBP. Although FBP expression is certainly one of the parameters affecting drug toxicity, taken alone it is not a sufficiently reliable predictor of cancer cell sensitivity to DDATHF.

Original languageEnglish
Pages (from-to)525-530
Number of pages6
JournalBritish Journal of Cancer
Volume73
Issue number4
Publication statusPublished - 1996

Fingerprint

Folic Acid
Carrier Proteins
Carcinoma
Cell Line
Membranes
Inhibitory Concentration 50
NIH 3T3 Cells
lometrexol
In Vitro Techniques
Hydroxymethyl and Formyl Transferases
Poisons
Type C Phospholipases
Phosphatidylinositols
Drug-Related Side Effects and Adverse Reactions
Neoplasms
Complementary DNA

Keywords

  • 5,10-dideazatetrahydrofolic acid
  • Folate-binding protein
  • Ovarian carcinoma

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Role of membrane folate-binding protein in the cytotoxicity of 5,10-dideazatetrahydrofolic acid in human ovarian carcinoma cell lines in vitro. / Sen, S.; Erba, E.; D'Incalci, M.; Bottero, F.; Canevari, S.; Tomassetti, A.

In: British Journal of Cancer, Vol. 73, No. 4, 1996, p. 525-530.

Research output: Contribution to journalArticle

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abstract = "Lometrexol (5,10-dideazatetrahydrofolic acid; DDATHF), is a specific inhibitor of glycinamideribonucleosyl (GAR) transformylase with anti-tumour activity in murine and human carcinomas. The cytotoxicity activity of DDATHF was evaluated in vitro in NIH/3T3 cells transfected with human α-folate-binding protein (FBP) complementary DNA to examine the role of the receptor. In FBP-transfecled NIH/3T3 (FBP-tNIH/3T3) cells, which internalised about three times more 5-methyltetrahydrofolic acid than the mock-transfected cells, the cytotoxic potential of DDATHF showed a clear increase. Subsequently, we analysed four ovarian carcinoma cell lines (OVCAR3, IGROV1, SKOV3 and SW626) expressing different amounts of FBP. Cells were conditioned to grow in medium depleted of folic acid then tested by MOv18 and folic acid binding. Only SKOV3 and SW626 cells grown in folic acid-depleted medium showed increased FBP expression, about 3-and 8-fold respectively. The cytotoxic potential of DDATHF was evaluated by a standard clonogenic assay. In a medium containing 2.27 μM folic acid the DDATHF IC50 values were 50 nM on OVCAR3, 500 nM on SW626 and 1000 nM on IGROV1. In folic acid-free medium IC50 values were 2 nM on OVCAR3 and SW626 and 40 nM on IGROV1. Only on SKOV3 cells was DDATHF cytotoxicity the same regardless of the amount of folic acid in the medium (IC50 8 nM). Thus, DDATHF did not inhibit the growth of IGROV1 cells depleted of folic acid after stripping FBP with phosphatidylinositol-phospholipase C, even at a dose toxic for cells constitutively expressing FBP. Although FBP expression is certainly one of the parameters affecting drug toxicity, taken alone it is not a sufficiently reliable predictor of cancer cell sensitivity to DDATHF.",
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AU - Sen, S.

AU - Erba, E.

AU - D'Incalci, M.

AU - Bottero, F.

AU - Canevari, S.

AU - Tomassetti, A.

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