Establishment and characterization of a cisplatin-resistant cell line, KB-R, derived from oral carcinoma cell line, KB.
Establishment and characterization of a cisplatin-resistant cell line, KB-R, derived from oral carcinoma cell line, KB.
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DOI：
10.3892/ijo.30.6.1325
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发表时间：
2007-06
影响因子：
5.2
通讯作者：
K. Negoro;Yukio Yamano;K. Fushimi;Kengo Saito;Ken Nakatani;M. Shiiba;H. Yokoe;H. Bukawa;K. Uzawa-K.
中科院分区：
文献类型：
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作者：
K. Negoro;Yukio Yamano;K. Fushimi;Kengo Saito;Ken Nakatani;M. Shiiba;H. Yokoe;H. Bukawa;K. Uzawa-K.
为探讨口腔癌细胞对顺铂（CDDP）耐药的机制，我们从口腔癌细胞系KB中筛选出了耐CDDP的细胞系KB-R。3-（3，4-二甲基噻唑-2-基）-2，5-二苯基四氮唑溴化物（MTT）测定表明KB-R对CDDP的抗性是KB的5.5倍。微阵列分析表明，与KB相比，KB-R中1，718个基因的表达水平至少提高了5倍或更多。采用半定量逆转录-聚合酶链反应（RT-PCR）和实时荧光定量PCR（real-time PCR）方法检测多药耐药基因中ATP结合盒（ABC）转运蛋白基因的表达情况。MRP 1和MRP 2表达上调，而MDR 1表达下调。使用Intimidity Pathway Analysis工具进行的通路和本体分析表明了三个高度显著的遗传网络，包括1，718个过表达基因中的105个，以及一个网络，包括35个“细胞间信号传导和相互作用”相关基因。我们的研究结果表明，这些细胞系，KB和KB-R可能是有用的，寻找候选基因负责CDDP耐药和进一步研究，以了解CDDP耐药的机制。
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.