Differential proteomic analysis of HL60 cells treated with secalonic acid F reveals caspase 3-induced cleavage of Rho GDP dissociation inhibitor 2.
Differential proteomic analysis of HL60 cells treated with secalonic acid F reveals caspase 3-induced cleavage of Rho GDP dissociation inhibitor 2.
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对用癸二酸 F 处理的 HL60 细胞进行差异蛋白质组分析，揭示了 caspase 3 诱导的 Rho GDP 解离抑制剂 2 裂解。
DOI：
10.3892/or.2012.2062
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发表时间：
2012
期刊：
影响因子：
4.2
通讯作者：
Xixiang Tang
中科院分区：
文献类型：
--
作者：
Ning Li;Zhiwei Yi;Yuqiao Wang;Qin Zhang;Tianhua Zhong;Yinkun Qiu;Zhen Wu;Xixiang Tang
二克隆酸F （SAF）已被发现，但其细胞毒活性和相关的细胞毒机制尚不清楚。本研究的目的是评价从深海真菌青霉菌sp. F11中分离的SAF对HL60细胞的细胞毒活性，并分析SAF对HL60细胞蛋白表达的影响。CCK-8实验和Annexin V-FLUOS/PI实验表明，SAF对HL60细胞增殖和诱导凋亡具有剂量依赖性和时间依赖性。4µg/ml SAF处理HL60细胞后，二维凝胶电泳（2-DE）分析显示，10个蛋白表达差异点（P<0.05）， 5个表达上调，5个表达下调。MALDI-TOF ms鉴定出3个位点（1下调，2上调）为Rho GDP解离抑制剂2 （RhoGDI 2）蛋白，Western blotting进一步证实全长RhoGDI 2的丰度降低，而caspase 3裂解产物的丰度增加。caspase 3抑制剂Ac-DEVD-CHO能抑制SAF的细胞毒作用，显著阻断RhoGDI 2的裂解。RhoGDI 2是Rho GTPase的胞质调节剂，其caspase 3裂解产物可促进细胞凋亡进程。我们的数据显示，SAF可能会调节HL60细胞中的RhoGDI 2水平，从而潜在地破坏对HL60细胞功能重要的细胞信号通路。
Secalonic acid F (SAF) has been previously identified, however, little is known about its cytotoxic activity and related cytotoxic mechanism. The aim of this study was to evaluate the cytotoxic activity of SAF isolated from a deep sea originated fungus Penicillium sp. F11 in HL60 cells and to analyze the differences in protein expression of HL60 cells treated with SAF. The CCK-8 assay and Annexin V-FLUOS/PI assay indicated that SAF displayed dose- and time-dependent inhibition of HL60 cell proliferation and induced apoptosis. Two-dimensional gel electrophoresis (2-DE) analysis of HL60 cells treated with SAF (4 µg/ml) revealed 10 differentially expressed protein spots (P<0.05), 5 upregulated and 5 downregulated. Three spots (1 downregulated and 2 upregulated) were identified as Rho GDP dissociation inhibitor 2 (RhoGDI 2) proteins by MALDI-TOF MS. Western blotting further demonstrated the decreased abundance of full-length RhoGDI 2 together with the increased abundance of caspase 3-cleaved product of RhoGDI 2. The caspase 3 inhibitor Ac-DEVD-CHO could suppress the cytotoxic effect of SAF and significantly block the cleavage of RhoGDI 2. RhoGDI 2 is a cytosolic regulator of Rho GTPase and the caspase 3-cleaved product of RhoGDI 2 can advance progression of the apoptotic process. Our data showed that SAF may modulate RhoGDI 2 levels in HL60 cells, thereby potentially disrupting cell signaling pathways important for HL60 cell function.
DOI：
10.1002/bdrb.20043
发表时间：
2005-06
期刊：
Birth defects research. Part B, Developmental and reproductive toxicology
影响因子：
--
作者：
V. Dhulipala;K. K. Maddali-K.;W. Welshons;C. Reddy
通讯作者：
V. Dhulipala;K. K. Maddali-K.;W. Welshons;C. Reddy
影响因子：
5.1
作者：
Gautschi, JT;Amagata, T;Crews, P
通讯作者：
Crews, P