Establishment and characterization of a cisplatin-resistant oral squamous cell carcinoma cell line, H-1R.
Cisplatin (CDDP) is a widely used potent chemotherapeutic agent for many malignancies. However, the mechanism of resistance to CDDP remains unclear. To investigate the molecular mechanism, we established a CDDP-resistant cell line (H-1R) from a CDDP-sensitive cell line (H-1) which was derived from moderately differentiated squamous cell carcinoma of the lower gingiva. The 3-(3,4-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay indicated that H-1R had a 10-fold greater resistance to CDDP than H-1. When we compared gene expression levels in the cell lines using an in-house cDNA microarray, which represented 2,201 genes originating from normal oral tissue, primary oral cancer, and oral cancer cell lines, 12 genes showing elevated mRNA expression in H-1R compared with H-1 were identified. Among them, the up-regulated expression of ATP-binding cassette transporter genes (MDR1, MRP1, and MRP2), CD55, and PGK1 and down-regulated expression of Caveolin 1 were further confirmed by semiquantitative reverse transcriptase-polymerase chain reaction (PCR) or real-time PCR. Our results suggest that H-1 and H-1R cell lines could be useful for elucidating the candidate genes responsible for CDDP resistance, including the genes found in this study.
Nakamura
Megumi
M
Department of Clinical Molecular Biology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.
Nakatani
Ken
K
Uzawa
Katsuhiro
K
Ono
Kanae
K
Uesugi
Hisako
H
Ogawara
Katsunori
K
Shiiba
Masashi
M
Bukawa
Hiroki
H
Yokoe
Hidetaka
H
Wada
Takeshi
T
Fujita
Shigeyuki
S
Tanzawa
Hideki
H
D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis / Q000235:genetics; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D002945:Cisplatin / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D009062:Mouth Neoplasms / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D014407:Tumor Cells, Cultured
D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D002945:Cisplatin