实时荧光定量PCR法检测人骨肉瘤耐MTX细胞系中RFC、DHFR、GST-π的mRNA表达Detection of RFC,DHFR and GST-π expression in human osteosarcoma cell line with MTX-resistence by FQ-PCR
于明东;李书忠;
摘要(Abstract):
[目的]研究RFC(还原叶酸载体)、GST-π(谷胱甘肽S转移酶π)、DHFR(二氢叶酸还原酶)mRNA在人骨肉瘤U2-OS细胞系及人骨肉瘤耐药细胞系U2-OS/R1-R3中的表达差异,并探讨其在人骨肉瘤MTX化疗耐药中的意义。[方法]采用冲击并逐步增加诱导药物浓度诱导的方法,诱导人骨肉瘤细胞系U2-OS,并建立耐MTX细胞系(U2-OS/R1-R3),利用荧光定量PCR技术在mRNA水平检测RFC、GST-π、DHFR在U2-OS及(U2-OS/R1-R3)中的表达。[结果]本试验成功建立三株人骨肉瘤耐药细胞系U2-OS/R1-R3中,荧光定量PCR结果显示MTX耐药与DHFRmRNA表达增加有关,与RFCmRNA的表达降低有关,与GST-πmRNA的表达增加有关。[结论]本试验在基因水平探讨人骨肉瘤细胞MTX化疗耐药机制,DHFR、GST-π基因表达的增加及RFC基因表达的降低参与了人骨肉瘤细胞的MTX耐药,为临床探索骨肉瘤患者的MTX耐药机制提供了重要的依据,为临床筛选MTX化疗不敏感患者提供重要的依据。
关键词(KeyWords): 实时荧光定量PCR;甲氨喋呤(MTX)还原叶酸载体(RFC);二氢叶酸还原酶(DHFR);谷胱甘肽S转移酶(GST-π)
基金项目(Foundation):
作者(Author): 于明东;李书忠;
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参考文献(References):
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