Down-regulation of argininosuccinate synthetase is associated with cisplatin resistance in hepatocellular carcinoma cell lines: implications for PEGylated arginine deiminase combination therapy.
Down-regulation of argininosuccinate synthetase is associated with cisplatin resistance in hepatocellular carcinoma cell lines: implications for PEGylated arginine deiminase combination therapy.
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DOI：
10.1186/1471-2407-14-621
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发表时间：
2014-08-28
期刊：
影响因子：
3.8
通讯作者：
Thomson JA
中科院分区：
文献类型：
--
作者：
McAlpine JA;Lu HT;Wu KC;Knowles SK;Thomson JA
许多晚期人类肿瘤，包括肝细胞癌（HCC），由于精氨酸琥珀酸合成酶（ASS1）的下调而对精氨酸产生营养不良，ASS1是精氨酸合成的限速酶。精氨酸降低剂聚乙二醇化精氨酸脱亚胺酶（ADI-PEG 20）在临床试验中显示出治疗精氨酸营养不良肿瘤的单药疗效，目前正在评估与顺铂联合治疗其他类型癌症的疗效。通过ASS1启动子甲基化的表观遗传沉默已经在其他类型的癌症中得到证实，并且在卵巢癌中也观察到ASS1表达与顺铂耐药之间的相互关系。然而，在HCC中，ASS1下调的机制以及与顺铂耐药的相关性尚未探讨。本研究探讨了ADI-PEG 20和顺铂敏感性与HCC中ASS1表达的关系。此外，我们展示了该生物标志物如何被顺铂单独和与ADI-PEG 20联合调节。western blot检测ASS1蛋白在未处理和药物处理的人HCC细胞系中的表达。利用基于发光的细胞活力测定法建立了这些细胞系中ASS1蛋白水平、ADI-PEG 20敏感性和顺铂耐药性之间的相关性。通过亚硫酸盐转化和甲基化特异性PCR分析ASS1的表观遗传调控。在HCC细胞系中，ASS1蛋白表达缺失、ASS1启动子甲基化、对ADI-PEG 20的敏感性与顺铂耐药性之间存在良好的相关性。此外，顺铂治疗可下调肝癌细胞系中ASS1蛋白的表达。然而，在临床相关浓度下，ADI-PEG 20和顺铂联合使用可以恢复大多数细胞系的ASS1蛋白水平。HCC细胞系中ASS1沉默与顺铂耐药和ADI-PEG 20敏感性同时相关，这表明HCC治疗的联合治疗策略很有前景。
Many advanced human tumors, including hepatocellular carcinomas (HCC) are auxotrophic for arginine due to down-regulation of argininosuccinate synthetase (ASS1), the rate-limiting enzyme in arginine synthesis. The arginine-lowering agent PEGylated arginine deiminase (ADI-PEG 20) has shown efficacy as a monotherapy in clinical trials for treating arginine-auxotrophic tumors and is currently being evaluated in combination with cisplatin in other cancer types. Epigenetic silencing via methylation of the ASS1 promoter has been previously demonstrated in other cancer types, and a reciprocal relationship between ASS1 expression and cisplatin resistance has also been observed in ovarian cancer. However, the mechanism of ASS1 down-regulation, as well as the correlation with cisplatin resistance has not been explored in HCC. The present study investigates ADI-PEG 20 and cisplatin sensitivities in relation to ASS1 expression in HCC. In addition, we show how this biomarker is regulated by cisplatin alone and in combination with ADI-PEG 20. ASS1 protein expression in both untreated and drug treated human HCC cell lines was assessed by western blot. The correlation between ASS1 protein levels, ADI-PEG 20 sensitivity and cisplatin resistance in these cell lines was established using a luminescence-based cell viability assay. Epigenetic regulation of ASS1 was analyzed by bisulfite conversion and methylation-specific PCR. A good correlation between absence of ASS1 protein expression, ASS1 promoter methylation, sensitivity to ADI-PEG 20 and resistance to cisplatin in HCC cell lines was observed. In addition, cisplatin treatment down-regulated ASS1 protein expression in select HCC cell lines. While, at clinically relevant concentrations, the combination of ADI-PEG 20 and cisplatin restored ASS1 protein levels in most of the cell lines studied. ASS1 silencing in HCC cell lines is associated with simultaneous cisplatin resistance and ADI-PEG 20 sensitivity which suggests a promising combination therapeutic strategy for the management of HCC.
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影响因子：
11.2
作者：
Kim, Randie H.;Coates, Jodi M.;Bowles, Tawnya L.;McNerney, Gregory P.;Sutcliffe, Julie;Jung, Jae U.;Gandour-Edwards, Regina;Chuang, Frank Y. S.;Bold, Richard J.;Kung, Hsing Jien
通讯作者：
Kung, Hsing Jien
影响因子：
8.8
作者：
通讯作者：
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影响因子：
45.3
作者：
Izzo, F;Marra, P;Curley, SA
通讯作者：
Curley, SA
影响因子：
6.2
作者：
Dillon, BJ;Prieto, VG;Clark, MA
通讯作者：
Clark, MA
影响因子：
45.3
作者：
Ascierto, PA;Scala, S;Logan, TF
通讯作者：
Logan, TF