Involvement of NF-κB activation in the cisplatin resistance of human epidermoid carcinoma KCP-4 cells
Involvement of NF-κB activation in the cisplatin resistance of human epidermoid carcinoma KCP-4 cells
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DOI：
10.3892/or.2012.1801
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发表时间：
2012-07-01
期刊：
影响因子：
4.2
通讯作者：
Kariyazono, Hiroko
中科院分区：
文献类型：
--
作者：
Oiso, Shigeru;Ikeda, Ryuji;Kariyazono, Hiroko
顺铂是目前治疗多种癌症最有效的抗癌药物之一。尽管它具有治疗作用，但在持续治疗下获得的内在抵抗力限制了它在癌症治疗中的好处。KCP-4是由人表皮样癌KB-3-1细胞分化而来的顺铂耐药细胞株。由于外排泵的存在显著减少顺铂在KCP-4细胞中的蓄积，该泵被认为与KCP-4细胞对顺铂的耐药性有关。然而，鉴于KCP-4细胞与KB-3-1细胞相比对顺铂具有巨大的耐药性，有可能存在另一种机制。本研究的目的是探讨核因子-kappaB的激活是否参与了KCP-4细胞对顺铂的耐药。我们用免疫印迹法检测核转录因子-kappaB移位到核内的水平作为核因子-kappaB活化的指标。KCP-4细胞中核因子-kappaB的活化水平高于KB-3-I细胞。KCP-4细胞用顺铂和姜黄素联合处理，姜黄素是一种核因子-kappa B的激活抑制剂，随后用3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium溴化物的四甲基偶氮唑盐比色法测定细胞的存活率。在10 mU/L姜黄素存在下，KCP-4细胞对100和300 mU/L顺铂的敏感性增加。此外，姜黄素还降低了KCP-4细胞中NF-kappa B的活化水平，并抑制了受NF-kappa B调控的凋亡相关蛋白Bcl2、Bclxl和Survivin的表达水平。我们的结果表明，KCP-4细胞与KB-3-1细胞相比，对顺铂的高耐药性是由多种机制造成的，而不是增加顺铂外流，包括激活NF-kappa B。
cis-Diamminedichloroplatinum II (cisplatin) is one of the most potent antitumor agents for the treatment of various types of cancer. In spite of its therapeutic usefulness, the intrinsic resistance acquired under continuous treatment limits its benefit in cancer therapy. KCP-4, a cisplatin-resistant cell line, was derived from human epidermoid carcinoma KB-3-1 cells. Since the accumulation of cisplatin in KCP-4 cells is markedly reduced by the presence of an efflux pump, this pump is thought to be related to cisplatin resistance of the KCP-4 cells. However, given that KCP-4 cells are tremendously resistant to cisplatin compared with KB-3-1 cells, it is possible that another mechanism exists. The aim of this study was to investigate whether the activation of nuclear factor-kappa B (NF-kappa B) contributes to the cisplatin resistance of KCP-4 cells. We used the level of translocated NF-kappa B into the nucleus, determined by immunoblot analysis, as the indicator of NF-kappa B activation. The activation level of NF-kappa B was higher in KCP-4 cells than in KB-3-I cells. KCP-4 cells were treated with a combination of cisplatin and curcumin, an inhibitor of NF-kappa B activation, and the cell viabilities were subsequently determined by the MTT assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. In the presence of 10 mu mol/l curcumin, we found that the sensitivity of KCP-4 cells to 100 and 300 mu mol/l cisplatin was augmented. Additionally, curcumin reduced the activation levels of NF-kappa B in KCP-4 cells, and suppressed the expression levels of Bcl-2, Bcl-xL and survivin, which are apoptosis-related proteins regulated by NF-kappa B. Our results suggest that the high cisplatin resistance of KCP-4 cells compared with KB-3-1 cells results from multiple mechanisms other than increased cisplatin efflux, including the activation of NF-kappa B.