Inhibition of PARP Activity by PJ-34 Leads to Growth Impairment and Cell Death Associated With Aberrant Mitotic Pattern and Nucleolar Actin Accumulation in M14 Melanoma Cell Line
Inhibition of PARP Activity by PJ-34 Leads to Growth Impairment and Cell Death Associated With Aberrant Mitotic Pattern and Nucleolar Actin Accumulation in M14 Melanoma Cell Line
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DOI：
10.1002/jcp.21964
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发表时间：
2010-02-01
影响因子：
5.6
通讯作者：
Caiafa, Paola
中科院分区：
文献类型：
--
作者：
Chevanne, Marta;Zampieri, Michele;Caiafa, Paola
PARP活性抑制剂阻止DNA损伤恢复的能力表明这些药物可用作癌症治疗的化学和放射增敏剂。我们在培养的人M14黑色素瘤细胞上进行的研究旨在检查PJ-34，一种有效的第二代PARP活性抑制剂，本身是否能够在没有任何DNA损伤剂的情况下影响这些癌细胞的活力。使用延时视频显微镜，我们证明，10 μ M PJ-34治疗诱导严重的有丝分裂缺陷，导致细胞增殖和细胞死亡的显着减少。PJ-34的细胞毒性作用通过细胞活力分析和克隆形成测定进一步证实。缺乏典型的细胞凋亡标志物使我们能够排除这种细胞死亡。未发现单链和/或双链DNA损伤。免疫荧光分析显示，在几个细胞和随后的多核异常有丝分裂的情况下，细胞死亡的非典型方式：有丝分裂灾难。核仁内聚合肌动蛋白异常积累的检测是值得注意的。总之，我们的结果表明，通过PJ-34靶向PARP活性，癌细胞存活受到独立于DNA损伤修复的影响。有两个发现值得注意：（a）如果随后用10 μ M PJ-34处理24小时以获得相同的细胞毒性作用，则顺铂浓度可降低四分之三;（B）依赖于PJ-34处理的作用是可逆的。我们的数据表明，为了减少顺铂化疗期间对非肿瘤细胞的伤害，后者可以与PJ-34治疗相结合。J.细胞。222：401-410，2010。(C)2009威利-利斯公司
The capability of PARP activity inhibitors to prevent DNA damage recovery suggested the use of these drugs as chemo- and radio-sensitisers for cancer therapy. Our research, carried out on cultured human M 14 melanoma cells, was aimed to examine if PJ-34, a potent PARP activity inhibitor of second generation, was per se able to affect the viability of these cancer cells without any DNA damaging agents. Using time-lapse videomicroscopy, we evidenced that 10 mu M PJ-34 treatment induced severe mitotic defects leading to dramatic reduction of cell proliferation and to cell death. PJ-34 cytotoxic effect was further confirmed by analysis of cell viability and clonogenic assay. Absence of canonic apoptosis markers allowed us to exclude this kind of cell death. No single and/or double stranded DNA damage was evidenced. Immunofluorescence analysis showed an aberrant mitotic scenario in several cells and subsequent multinucleation suggesting an atypical way for cells to die: the mitotic catastrophe. The detection of aberrant accumulation of polymerised actin inside the nucleolus was noteworthy. Taken together, our results demonstrate that, targeting PARP activity by PJ-34, cancer cell survival is affected independently of DNA damage repair. Two findings are remarkable: (a) cisplatin concentration can be reduced by three quarters if it is followed by treatment with 10 mu M PJ-34 for 24 h to obtain the same citotoxic effect; (b) effects dependent on PJ-34 treatment are reversible. Our data suggest that, to reduce the harm done to non-tumour cells during chemotherapy with cisplatin, the latter could be coupled with PJ-34 treatment. J. Cell. Physiol. 222: 401-410, 2010. (C) 2009 Wiley-Liss, Inc.