Analysis of the medical treatment resistance factor in salivary gland carcinoma

唾液腺癌耐药因素分析

基本信息

批准号：
20791557
负责人：
DOTO Ryosuke
金额：
$ 2.75万
依托单位：
Matsumoto Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2009
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20791557/
关键词：
ABC transporter MDR1 MRP1 GST 抗癌剤多剤耐性薬剤排泄機構 MDRl MRPl

项目摘要

Objectives : The poor clinical response of salivary gland adenocarcinoma (SGA) to combined chemotherapy for head-and-neck cancer (HNC) suggests that the sensitivities and/or mechanisms of resistance to anti-cancer drugs differ between SGA and oral squamous cell carcinoma (SCC). The present study aimed to clarify whether expression of the ATP-binding cassette (ABC) transporters MDR1 and MRP1 is associated with multidrug resistance (MDR) in HNC, and to determine differences in the associated processes between SGA and SCC.Materials and Methods: The expression of ABC transporters and glutathione S-transferase enzymes was studied immunohistochemically in the normal salivary gland and in cancer tissues from patients. Expression of MDR1 and MRP1 was analyzed using Western blotting in both SGA and SCC cell lines following vincristine administration. We also determined mdr1 and mrp1 mRNA expression levels using reverse transcriptase-polymerase chain reaction (RT-PCR).Results : Immunohistochemical analysis revealed MDR1 and MRP1 expression in ductal cells of salivary glands, but not in the oral mucosal epithelium. When compared with SCC, SGA displayed expressed more intense MRP1 and more MDR1. Analysis in vitro indicated more MDR1, MRP1 and glutathione S-transferase (GST)-pi class enzyme expression in drug-resistant SGA cells than in parental SGA or drug-resistant and drug-sensitive SCC cell lines. Immunoblotting and PCR confirmed that the expression of GST-pi was increased relative to that of MRP1 in drug-resistant SGA cells. The MDR1 inhibitor verapamil obviously enhanced cytotoxicity in resistant SGA and SCC cells at low vincristine concentrations. The GST inhibitor curcumin also enhanced cytotoxicity, particularly in resistant SGA cells after the MDR1 efflux barrier had been reversed by verapamil.Conclusion : These results indicate that MRP1/GST-pi enzymes are involved in the acquired-resistance phenotype of SGA.

目的：唾液腺腺癌（SGA）对头颈癌（HNC）联合化疗的临床反应不佳表明，SGA 与口腔鳞状细胞癌之间抗癌药物的敏感性和/或耐药机制不同。 SC）。本研究旨在阐明 ATP 结合盒 (ABC) 转运蛋白 MDR1 和 MRP1 的表达是否与 HNC 中的多药耐药性 (MDR) 相关，并确定 SGA 和 SCC 之间相关过程的差异。通过免疫组织化学方法研究了正常唾液腺和患者癌症组织中 ABC 转运蛋白和谷胱甘肽 S-转移酶的表达。在施用长春新碱后，使用蛋白质印迹分析 SGA 和 SCC 细胞系中 MDR1 和 MRP1 的表达。我们还使用逆转录聚合酶链式反应 (RT-PCR) 测定了 mdr1 和 mrp1 mRNA 表达水平。结果：免疫组织化学分析显示 MDR1 和 MRP1 在唾液腺导管细胞中表达，但在口腔粘膜上皮中不表达。与SCC相比，SGA表现出表达更强烈的MRP1和更多的MDR1。体外分析表明，耐药 SGA 细胞中 MDR1、MRP1 和谷胱甘肽 S-转移酶 (GST)-pi 类酶的表达高于亲本 SGA 或耐药和药物敏感 SCC 细胞系。免疫印迹和PCR证实耐药SGA细胞中GST-pi的表达相对于MRP1有所增加。 MDR1抑制剂维拉帕米在低长春新碱浓度下明显增强耐药SGA和SCC细胞的细胞毒性。 GST 抑制剂姜黄素也增强了细胞毒性，特别是在维拉帕米逆转 MDR1 流出屏障后的耐药 SGA 细胞中。结论：这些结果表明 MRP1/GST-pi 酶参与了 SGA 的获得性耐药表型。