Atypical multidrug resistance: Breast cancer resistance protein messenger RNA expression in mitroxantrone-selected cell lines
Atypical multidrug resistance: Breast cancer resistance protein messenger RNA expression in mitroxantrone-selected cell lines
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DOI：
10.1093/jnci/91.5.429
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发表时间：
1999-03-03
期刊：
影响因子：
--
通讯作者：
Doyle, LA
中科院分区：
文献类型：
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作者：
Ross, DD;Yang, WD;Doyle, LA
背景：在细胞毒剂米托蒽醌存在下生长的人癌细胞株经常发生耐药，与减少细胞内药物积累有关，而不增加已知耐药转运体P-糖蛋白和多药耐药蛋白(也称为多药耐药相关蛋白)的表达。乳腺癌耐药蛋白(BCRP)是一种新近发现的三磷酸腺苷结合盒转运蛋白，与米托蒽醌和蒽环类药物耐药有关。这项研究是为了测试在米托蒽醌存在的情况下，选择生长的细胞系中BCRP过度表达的流行率。方法：从亲本细胞和药物筛选细胞中提取细胞总RNA或聚A(+)RNA和基因组DNA。分别用Northern杂交和Southern印迹杂交分析BCRP信使RNA的表达和BCRP基因的扩增。结果：经米托蒽醌筛选后获得的多种耐药细胞系，包括人乳腺癌(MCF-7)、结肠癌(S1和HT29)、胃癌(EPG85-257)、纤维肉瘤(EPF86-079)和骨髓瘤(8226)等多种耐药细胞系，BCRP基因在这些细胞中也得到了扩增。结论：在米托蒽醌筛选的多药耐药细胞系中，经常观察到BCRP mRNA的过度表达，提示BCRP可能是该药物诱导的主要细胞防御机制。BCRP很可能是推测存在于这些细胞系中的“米托蒽醌转运体”。
Background: Human cancer cell lines grown in the presence of the cytotoxic agent mitoxantrone frequently develop resistance associated with a reduction in intracellular drug accumulation without increased expression of the known drug resistance transporters P-glycoprotein and multidrug resistance protein (also known as multidrug resistance-associated protein). Breast cancer resistance protein (BCRP) is a recently described adenosine triphosphate-binding cassette transporter associated with resistance to mitoxantrone and anthracyclines. This study was undertaken to test the prevalence of BCRP overexpression in cell lines selected for growth in the presence of mitoxantrone. Methods: Total cellular RNA or poly A(+) RNA and genomic DNA were isolated from parental and drug-selected cell fines. Expression of BCRP messenger RNA (mRNA) and amplification of the BCRP gene were analyzed by northern and Southern blot hybridization, respectively, Results: A variety of drug-resistant human cancer cell lines derived by selection with mitoxantrone markedly overexpressed BCRP mRNA; these cell lines included sublines of human breast carcinoma (MCF-7), colon carcinoma (S1 and HT29), gastric carcinoma (EPG85-257), fibrosarcoma (EPF86-079), and myeloma (8226) origins, Analysis of genomic DNA from BCRP-overexpressing MCF-7/MX cells demonstrated that the BCRP gene was also amplified in these cells. Conclusions: Overexpression of BCRP mRNA is frequently observed in multidrug-resistant cell lines selected with mitoxantrone, suggesting that BCRP is likely to be a major cellular defense mechanism elicited in response to exposure to this drug. It is likely that BCRP is the putative "mitoxantrone transporter" hypothesized to be present in these cell lines.