Cyclin B1 Overexpression Induces Cell Death Independent of Mitotic Arrest
Cyclin B1 Overexpression Induces Cell Death Independent of Mitotic Arrest
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DOI：
10.1371/journal.pone.0113283
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发表时间：
2014-11-21
期刊：
影响因子：
3.7
通讯作者：
Chambers, Timothy C.
中科院分区：
文献类型：
--
作者：
Eichhorn, Joshua M.;Kothari, Anisha;Chambers, Timothy C.
微管抑制剂广泛应用于癌症化疗。这些药物的特征是诱导有丝分裂停滞和细胞死亡，但两者之间的联系机制尚未确定。问题之一是癌细胞对这些试剂的敏感性差异很大，因此很难比较来自不同系统的数据。为了缓解这个问题，我们试图通过在HeLa细胞中表达不可降解的细胞周期蛋白B（R42 A）来分子诱导有丝分裂死亡并研究其机制。然而，这种方法未能诱导显着的有丝分裂停滞，Cdk 1激活，或抗凋亡Bcl-2蛋白的磷酸化，所有的特征与微管抑制剂处理的细胞。此外，cyclin B1-R42 A诱导细胞快速死亡，当在同步化细胞中表达时，细胞死亡发生在G1期。降低质粒浓度降低转染效率，但恢复有丝分裂停滞和消除非特异性死亡。这些结果表明，细胞周期蛋白B1的不适当的过度表达导致非特异性细胞死亡，并建议谨慎使用它的有丝分裂事件的研究。
Microtubule inhibitors are widely used in cancer chemotherapy. These drugs characteristically induce mitotic arrest and cell death but the mechanisms linking the two are not firmly established. One of the problems is that cancer cells vary widely in their sensitivity to these agents, and thus comparison of data from different systems is difficult. To alleviate this problem we sought to molecularly induce mitotic death and study its mechanisms, by expressing non-degradable cyclin B (R42A) in HeLa cells. However, this approach failed to induce significant mitotic arrest, Cdk1 activation, or phosphorylation of anti-apoptotic Bcl-2 proteins, all characteristics of cells treated with microtubule inhibitors. Furthermore, cyclin B1-R42A induced rapid cell death, and when expressed in synchronized cells, cell death occurred in G1 phase. Decreasing the plasmid concentration reduced transfection efficiency but restored mitotic arrest and eliminated non-specific death. These results show that inappropriate overexpression of cyclin B1 causes non-specific cell death and suggest caution in its use for the study of mitotic events.