Microtubule S-glutathionylation as a potential approach for antimitotic agents.
Microtubule S-glutathionylation as a potential approach for antimitotic agents.
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DOI：
10.1186/1471-2407-12-245
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发表时间：
2012-06-15
期刊：
影响因子：
3.8
通讯作者：
Guan X
中科院分区：
文献类型：
--
作者：
Chen W;Seefeldt T;Young A;Zhang X;Zhao Y;Ruffolo J;Kaushik RS;Guan X
微管已成为抗癌药物开发最有效的靶标之一。用这些药物治疗的癌细胞的特点是细胞停滞在 G2/M 期。因此，微管靶向药物被称为抗有丝分裂剂。然而，当前抗有丝分裂药物的临床应用受到新出现的耐药性的阻碍，这是癌症治疗失败的主要原因。抗有丝分裂药物的临床成功和新出现的耐药性促使人们寻找新的抗有丝分裂药物，特别是具有新作用机制的抗有丝分裂药物。本研究的目的是确定癌细胞中的微管是否可以被S-谷胱甘肽化，以及谷胱甘肽化是否会导致微管功能障碍和细胞生长抑制。该研究将确定微管 S-谷胱甘肽化是否可以成为抗有丝分裂剂的新方法。 2-乙酰氨基-3-[4-(2-乙酰氨基-2-羧乙基硫基羰基氨基)苯基氨基甲酰基硫基]丙酸(2-AAPA)用作诱导微管S-谷胱甘肽化的工具。 UACC-62 细胞（一种人类黑色素瘤细胞系）被用作癌细胞模型。采用谷胱甘肽 S-转移酶 (GST)-琼脂糖珠进行下拉测定，然后进行蛋白质印迹分析来确认微管 S-谷胱甘肽化。使用小鼠单克隆抗α-微管蛋白-FITC进行免疫荧光显微镜研究S-谷胱甘肽化对微管功能的影响；主要是聚合和解聚。采用流式细胞术检测S-谷胱甘肽化对细胞周期分布和细胞凋亡的影响。通过使用 Zeiss AXIO Observer A1 显微镜跟踪细胞形态变化。使用 10 种人类癌细胞系研究了 2-AAPA 对癌细胞生长的抑制作用。我们的研究表明，在 2-AAPA 存在的情况下，UACC-62 细胞的细胞形态发生了变化，微管发生了 S-谷胱甘肽化。因此，发现微管解聚并且细胞停滞在G2/M期。发现受影响的细胞发生凋亡。癌症生长抑制实验表明，对微管产生作用所需的 2-AAPA 浓度与产生癌细胞生长抑制的浓度相一致。这项研究的数据证实，微管 S-谷胱甘肽化会导致微管功能障碍和细胞生长抑制，并且可以成为开发抗有丝分裂剂的新方法。
Microtubules have been one of the most effective targets for the development of anticancer agents. Cancer cells treated by these agents are characterized by cell arrest at G2/M phase. Microtubule-targeting drugs are, therefore, referred to as antimitotic agents. However, the clinical application of the current antimitotic drugs is hampered by emerging drug resistance which is the major cause of cancer treatment failure. The clinical success of antimitotic drugs and emerging drug resistance has prompted a search for new antimitotic agents, especially those with novel mechanisms of action. The aim of this study was to determine whether microtubules can be S-glutathionylated in cancer cells and whether the glutathionylation will lead to microtubule dysfunction and cell growth inhibition. The study will determine whether microtubule S-glutathionylation can be a novel approach for antimitotic agents. 2-Acetylamino-3-[4-(2-acetylamino-2-carboxyethylsulfanylcarbonylamino)phenyl carbamoylsulfanyl]propionic acid (2-AAPA) was used as a tool to induce microtubule S-glutathionylation. UACC-62 cells, a human melanoma cell line, were used as a cancer cell model. A pull-down assay with glutathione S-transferase (GST)-agarose beads followed by Western blot analysis was employed to confirm microtubule S-glutathionylation. Immunofluorescence microscopy using a mouse monoclonal anti-α-tubulin-FITC was used to study the effect of the S-glutathionylation on microtubule function; mainly polymerization and depolymerization. Flow cytometry was employed to examine the effect of the S-glutathionylation on cell cycle distribution and apoptosis. Cell morphological change was followed through the use of a Zeiss AXIO Observer A1 microscope. Cancer cell growth inhibition by 2-AAPA was investigated with ten human cancer cell lines. Our investigation demonstrated that cell morphology was changed and microtubules were S-glutathionylated in the presence of 2-AAPA in UACC-62 cells. Accordingly, microtubules were found depolymerized and cells were arrested at G2/M phase. The affected cells were found to undergo apoptosis. Cancer growth inhibition experiments demonstrated that the concentrations of 2-AAPA required to produce the effects on microtubules were compatible to the concentrations producing cancer cell growth inhibition. The data from this investigation confirms that microtubule S-glutathionylation leads to microtubule dysfunction and cell growth inhibition and can be a novel approach for developing antimitotic agents.
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DOI：
10.1016/j.jpba.2008.08.033
发表时间：
2008-12-15
影响因子：
3.4
作者：
Chen, Wei;Zhao, Yong;Seefeldt, Teresa;Guan, Xiangming
通讯作者：
Guan, Xiangming
影响因子：
4.8
作者：
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4.8
作者：
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通讯作者：
Murphy, MP
DOI：
10.1016/j.bbrc.2003.12.126
发表时间：
2004-02-06
影响因子：
3.1
作者：
Landino, LM;Moynihan, KL;Kennett, KL
通讯作者：
Kennett, KL
影响因子：
4.1
作者：
Kaur, Gurmeet;Hollingshead, Melinda;Agarwal, Seema
通讯作者：
Agarwal, Seema