Establishment of cisplatin-resistant ovarian yolk sac tumor cells and investigation of the mechanism of cisplatin resistance using this cell line
Establishment of cisplatin-resistant ovarian yolk sac tumor cells and investigation of the mechanism of cisplatin resistance using this cell line
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顺铂耐药卵巢卵黄囊肿瘤细胞的建立及顺铂耐药机制研究
DOI：
10.1159/000320744
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
他
中科院分区：
文献类型：
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作者：
Shibata K;Kajiyama H;他
背景：顺铂是治疗卵巢卵黄囊瘤（YST）的关键药物，但有复发的可能。负责顺铂resistance.Methods的分子机制的细节：我们建立了顺铂耐药的卵巢YST细胞（NOY 1-CR）从父母NOY 1。为了表征这些细胞，我们检查了对其他抗癌药物的交叉耐药性。然后，进行cDNA微阵列分析以定量NOY 1和NOY 1-CR细胞中的基因表达。采用实时荧光定量PCR和Western blotting方法比较了几个与耐药相关的潜在基因的表达情况。结果：NOY 1-CR细胞对顺铂的IC 50是亲本细胞的22.3倍。NOY 1-CR细胞对某些药物有交叉耐药，但对VP-16和博来霉素无交叉耐药。微阵列分析在NOY 1-CR细胞中鉴定出315个上调基因和412个下调基因。通过GSTA 1 siRNA敲低在耐药细胞中上调的GSTA 1，恢复了NOY 1-CR细胞对顺铂的敏感性。结论：我们的数据提示了顺铂耐药的分子机制，并显示GSTA 1有可能成为顺铂耐药卵巢YST的一个新的治疗靶点。
Background:Cisplatin is used as a key drug for ovarian yolk sac tumor (YST), but relapse may occur. Details of the molecular mechanism responsible for cisplatin resistance remain unclear.Methods:We established cisplatin-resistant ovarian YST cells (NOY1-CR) from parent NOY1. To characterize these cells, we examined cross-resistance to other anticancer drugs. Then, cDNA microarray analysis was performed to quantify gene expression in NOY1 and NOY1-CR cells. The expression of several potential genes related to drug resistance was compared with parent cells by real-time PCR and Western blotting. Knockdown experiments using small interfering RNA (siRNA) were also performed to confirm the genetic association with drug resistance.Results:The IC50for cisplatin of NOY1-CR was 22.3-fold higher than that for parent cells. NOY1-CR cells showed cross-resistance to some drugs, but not to VP-16 and bleomycin. Microarray analysis identified 315 up-regulated and 412 down-regulated genes in NOY1-CR cells. Knockdown of GSTA1, which was up-regulated in resistant cells, by GSTA1 siRNA restored cisplatin sensitivity in NOY1-CR cells.Conclusions:Our data suggest the molecular mechanisms of cisplatin resistance and show the potential for GSTA1 to become a novel therapeutic target for cisplatin-resistant ovarian YST.