Loss of p53 function confers high-level multidrug resistance in neuroblastoma cell lines.
Loss of p53 function confers high-level multidrug resistance in neuroblastoma cell lines.
复制标题
DOI：
--
复制
发表时间：
2001-08
期刊：
影响因子：
11.2
通讯作者：
N. Keshelava;J. Zuo;Ping Chen;Sitara N. Waidyaratne;Marian Luna;C. Gomer;T. Triche;C. Reynolds
中科院分区：
文献类型：
--
作者：
N. Keshelava;J. Zuo;Ping Chen;Sitara N. Waidyaratne;Marian Luna;C. Gomer;T. Triche;C. Reynolds
神经母细胞瘤在化疗期间，特别是清髓性放化疗期间可以获得持续的高水平耐药性。 p53 突变在原发性神经母细胞瘤中很少见，但 p53 功能的丧失可能在多药耐药性中发挥作用。我们通过测量 7 个 L-PAM 敏感和 11 个 L-PAM 抗性神经母细胞瘤细胞系中对马法兰 (L-PAM) 的 p21 和/或 MDM2 蛋白诱导来确定 p53 功能。 p53 在 7/7 药物敏感细胞系中发挥作用，但仅在 4/11 耐药细胞系中发挥作用 (P = 0.01)。在缺乏 p53 功能的 7 个细胞系中，有 4 个通过微阵列 GeneChip p53 检测和自动测序检测到了 p53 突变，而具有功能性 p53 的 6 个细胞系没有 p53 突变的证据。所有具有野生型 (wt) p53 的细胞系都显示出 p53-HBS/CAT 报告基因的强反式激活，而具有突变型 p53 的四种细胞系未能反式激活 p53 HBS/CAT。在具有 wt p53 的两个 p53 无功能细胞系中观察到 MDM2 蛋白过度表达（相对于 p53 功能细胞系）；其中一项显示 MDM2 的基因组扩增。在耐药细胞系中检测到非功能性和突变的 p53，而治疗前源自同一患者的敏感细胞系具有功能性和野生型 p53。通过将人乳头瘤病毒 16 E6（降解 p53）转导至两种具有完整 p53 的药物敏感神经母细胞瘤细胞系中，选择性地丧失 p53 功能，从而导致对 L-PAM、卡铂和依托泊苷的高水平耐药性。使用神经母细胞瘤细胞系获得的这些数据表明，在一些复发性神经母细胞瘤中观察到的高水平耐药性可归因于 p53 突变和/或化疗期间获得的 p53 功能丧失。如果在患者肿瘤样本中得到证实，这些数据支持开发独立于 p53 的疗法，以巩固和/或挽救复发性神经母细胞瘤。
Neuroblastomas can acquire a sustained high-level drug resistance during chemotherapy and especially myeloablative chemoradiotherapy. p53 mutations are rare in primary neuroblastomas, but a loss of p53 function could play a role in multidrug resistance. We determined p53 function by measuring induction of p21 and/or MDM2 proteins in response to melphalan (L-PAM) in seven L-PAM-sensitive and 11 L-PAM-resistant neuroblastoma cell lines. p53 was functional in seven/seven drug-sensitive but in only 4/11 drug-resistant cell lines (P = 0.01). In four of the seven cell lines lacking p53 function, mutations of p53 were detected by the microarray GeneChip p53 Assay and automated sequencing, whereas six cell lines with functional p53 had no evidence of p53 mutations. All of the cell lines with wild-type (wt) p53 showed a strong transactivation of the p53-HBS/CAT reporter gene, whereas the four cell lines with mutant p53 failed to transactivate p53 HBS/CAT. Overexpression of MDM2 protein (relative to p53 functional lines) was seen in two p53-nonfunctional cell lines with wt p53; one showed genomic amplification of MDM2. Nonfunctional and mutated p53 was detected in a resistant cell line, whereas a sensitive cell line derived from the same patient before treatment had functional and wt p53. Loss of p53 function was selectively achieved by transduction of human papillomavirus 16 E6 (which degrades p53) into two drug-sensitive neuroblastoma cell lines with intact p53, causing high-level drug resistance to L-PAM, carboplatin, and etoposide. These data obtained with neuroblastoma cell lines suggest that the high-level drug resistance observed in some recurrent neuroblastomas is attributable to p53 mutations and/or a loss of p53 function acquired during chemotherapy. If confirmed in patient tumor samples, these data support development of p53-independent therapies for consolidation and/or salvage of recurrent neuroblastomas.