Establishment and characterization of a cisplatin-resistant cell line, KB-R, derived from oral carcinoma cell line, KB.
To investigate the mechanism of the resistance to cisplatin (CDDP), we established the CDDP-resistant cell line, KB-R, from CDDP-sensitive oral carcinoma cell line, KB. The 3-(3, 4-dimethyl-thiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) assay indicated that KB-R is 5.5-fold more resistant to CDDP than KB. Microarray analysis indicated that the expression levels of 1,718 genes were elevated at least five-fold or more in KB-R, compared with KB. The expression status of ATP binding cassette (ABC) transporter genes, which belong to multi-drug resistance genes, was confirmed by semiquantitative reverse transcriptase-polymerase chain reaction and real-time PCR. MRP1 and MRP2 were up-regulated, whereas MDR1 was down-regulated. Pathway and ontology analysis using the Ingenuity Pathway Analysis tool indicated three highly significant genetic networks including 105 of the 1,718 overexpressed genes and one network including 35 'cell-to-cell signaling and interaction' related genes. Our results suggested that these cell lines, KB and KB-R, may be useful for searching the candidate genes responsible for CDDP-resistance and for further study to understand the mechanism of CDDP-resistance.
Negoro
Kenji
K
Department of Oral and Maxillofacial Surgery, School of Medicine, Wakayama Medical University, Japan.
Yamano
Yukio
Y
Fushimi
Kazuaki
K
Saito
Kengo
K
Nakatani
Ken
K
Shiiba
Masashi
M
Yokoe
Hidetaka
H
Bukawa
Hiroki
H
Uzawa
Katsuhiro
K
Wada
Takeshi
T
Tanzawa
Hideki
H
Fujita
Shigeyuki
S
D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D019450:Genes, MDR; D006801:Humans; D007624:KB Cells; D009062:Mouth Neoplasms / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000970:Antineoplastic Agents; D002945:Cisplatin