Cyclosporin A, verapamil and S9788 reverse doxorubicin resistance in a human medullary thyroid carcinoma cell line.
Multidrug resistance was investigated in TT cells, a human medullary thyroid carcinoma (MTC) cell line and in normal thyrocytes. MDR1 mRNA was revealed by polymerase chain reaction (PCR) analysis both in normal and neoplastic cells despite the absence of glycoprotein P (Pgp) by immunohistochemistry using JSB-1 monoclonal antibody. Glutathione-S-transferase mRNA was undetectable by Northern blotting in TT cells. Doxorubicin-induced cytotoxicity was evaluated in TT cells with MTT, lacticodehydrogenase (LDH), glutathione (GSH) assays and neutral red uptake. IC50 values obtained for MTT assays were higher than those obtained with the three other tests. Cyclosporin A (CSA) (3 microM), verapamil (10 microM) and S9788 (5 microM) partially reversed the resistance to doxorubicin after a 48 h co-incubation (followed by a 24 h post-incubation for the S9788). Under these conditions, GSH levels were altered by verapamil and S9788, whereas CSA decreased LDH activity. CSA and verapamil had no effect on MTT assay. In conclusion this MTC cell line exhibited over-expression of the MDR1 gene and its resistance to doxorubicin can be partially reversed by CSA, verapamil and S9788.
Massart
C
C
Laboratoire de Biochimie A, CHU de Pontchaillou, Rennes, France.
Gibassier
J
J
Raoul
M
M
Pourquier
P
P
Leclech
G
G
Robert
J
J
Lucas
C
C
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis / Q000235:genetics*; D018276:Carcinoma, Medullary / Q000188:drug therapy* / Q000473:pathology; D016572:Cyclosporine / Q000494:pharmacology*; D004317:Doxorubicin / Q000494:pharmacology*; D018432:Drug Resistance, Multiple / Q000235:genetics; D004357:Drug Synergism; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics*; D010880:Piperidines / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000096:biosynthesis; D013964:Thyroid Neoplasms / Q000188:drug therapy* / Q000473:pathology; D014227:Triazines / Q000494:pharmacology*; D014700:Verapamil / Q000494:pharmacology*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D009363:Neoplasm Proteins; D010880:Piperidines; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014227:Triazines; C077022:S 9788; D004317:Doxorubicin; D016572:Cyclosporine; D014700:Verapamil; D005982:Glutathione Transferase; D005978:Glutathione