title: Evaluation of MDR1, LRP, MRP, and topoisomerase IIalpha gene mRNA transcripts before and after interferon-alpha, and correlation with the mRNA expression level of the telomerase subunits hTERT and TEP1 in five unselected human melanoma cell lines.
abstract: Intrinsic and acquired multidrug-resistance (MDR) and the activity of the enzyme telomerase have been demonstrated in human melanoma. A direct regulation of the MDR pathways and of telomerase by interpheron-alpha (IFN-alpha), which is currently used in the therapy of advanced cutaneous melanoma, has also been hypothesized. In this study, we used five melanoma cell lines not selected in vitro for drug resistance (Me665/2/21, Me665/2/60, HT-144, SK-MEL-28, and SK-MEL-5), which in a previous study, had shown different responses to IFN-alpha in terms of proliferation, apoptosis, telomerase activity and expression of mRNA for the human telomerase reverse transcriptase (hTERT). We investigated the expression of the multidrug resistance (MDR1) gene, multidrug resistance protein (MRP), lung resistance protein (LRP), topoisomerase IIalpha (Topo IIalpha), hTERT, and telomerase-associated protein (TEP1), which is shared by telomerase and vault MDR proteins at the mRNA expression level, using the reverse transcription-PCR (RT-PCR). All cell lines showed an intrinsic expression of hTERT, TEP1, and MDR gene transcripts (only MDR1 mRNA was under the detection level in SK-MEL-28 cells). After IFN-alpha exposure, we observed either no effect, a trend towards a decrease of hTERT, MRP, and Topo IIalpha, or an increase of TEP1, MDR1, and LRP mRNA expression in some cell lines. Effects were usually temporary and not always significant. No correlation was found between hTERT and TEP1 mRNA expression, whereas significant positive correlations were found between TEP1 and MDR1 mRNA, and between TEP1 and LRP mRNA. IFN-alpha modulates differently MDR gene transcripts in human melanoma cell lines. Positive correlation between TEP1 and LRP also seems to identify them as common targets of IFN-alpha effects.
authors.affiliation: Istituto di Anatomia e Istologia Patologica, Policlinico Le Scotte, Viale Bracci 6, I-53100 Siena, Italy. miracco@unisi.it
authors.forename: Clelia
authors.initials: C
authors.lastname: Miracco
authors.forename: Emilia
authors.initials: E
authors.lastname: Maellaro
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Pacenti
authors.forename: Barbara
authors.initials: B
authors.lastname: Del Bello
authors.forename: Marta A
authors.initials: MA
authors.lastname: Valentini
authors.forename: Pietro
authors.initials: P
authors.lastname: Rubegni
authors.forename: Luigi
authors.initials: L
authors.lastname: Pirtoli
authors.forename: Chiara
authors.initials: C
authors.lastname: Volpi
authors.forename: Rosa
authors.initials: R
authors.lastname: Santopietro
authors.forename: Piero
authors.initials: P
authors.lastname: Tosi
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis*; D000951:Antigens, Neoplasm; D002352:Carrier Proteins / Q000096:biosynthesis*; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis*; D004268:DNA-Binding Proteins; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use*; D008545:Melanoma / Q000378:metabolism; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000096:biosynthesis*; D013997:Time Factors; D020394:Vault Ribonucleoprotein Particles / Q000096:biosynthesis*; D000099187:Major Vault Protein
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000951:Antigens, Neoplasm; D002352:Carrier Proteins; D004250:DNA Topoisomerases, Type II; D004268:DNA-Binding Proteins; D016898:Interferon-alpha; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D019098:Telomerase; D020394:Vault Ribonucleoprotein Particles; D000099187:Major Vault Protein; C104342:TEP1 protein, human