Giri B, Gomes A, Debnath A, Saha A, Biswas A K, Dasgupta S C, Gomes A
Laboratory of Toxinology and Experimental Pharmacodynamics, Department of Physiology, University of Calcutta, 92, APC Road, Kolkata 700 009, India.
Toxicon. 2006 Sep 15;48(4):388-400. doi: 10.1016/j.toxicon.2006.06.011. Epub 2006 Jun 29.
The antiproliferative, cytotoxic and apoptogenic activities of Bufo melanostictus (Indian common toad) skin extract (TSE) on U937 and K562 leukemic cell line has been investigated. TSE significantly (P<0.001) reduced the time-dependent cell proliferation and decreased MTT values in U937 and K562 cells. TSE (IC50 doses) suppressed the proliferating cell nuclear antigen expression in both the cells. It was demonstrated that, TSE (IC50 doses) primarily arrested the U937 and K562 cells at G1 phase of the cell cycle. Confocal microscopy showed the altered fragmented nuclei and apoptotic bodies formation in TSE (IC50 doses) treated U937 and K562 cells. Membrane blebbing, cell surface shrinkage and perforation were observed through scanning electron microscope. TSE-induced DNA fragmentation in U937 and K562 cells was reflected in single-cell gel electrophoresis. TSE significantly (P<0.001) increase the length-width ratio of DNA mass as compared to control in comet assay. The flow cytometric analysis of annexin-V binding to the cancer cells further supported the apoptotogenic activity of TSE. The effect of TSE on normal human peripheral blood mononuclear cells viability and cytotoxicity was studied in culture and found to be less cytotoxic than on the U937 and K562 cells. The findings from the present study suggested that TSE might possess potent antineoplastic agent having antiproliferative, cytotoxic and apoptogenic activity against U937 and K562 myeloid leukemic cells.
已对黑眶蟾蜍（印度普通蟾蜍）皮肤提取物（TSE）对U937和K562白血病细胞系的抗增殖、细胞毒性和诱导凋亡活性进行了研究。TSE显著（P<0.001）降低了U937和K562细胞中时间依赖性的细胞增殖并降低了MTT值。TSE（IC50剂量）抑制了两种细胞中增殖细胞核抗原的表达。结果表明，TSE（IC50剂量）主要使U937和K562细胞停滞在细胞周期的G1期。共聚焦显微镜显示，经TSE（IC50剂量）处理的U937和K562细胞中出现了核碎片化改变和凋亡小体形成。通过扫描电子显微镜观察到膜泡形成、细胞表面收缩和穿孔。TSE诱导的U937和K562细胞DNA片段化在单细胞凝胶电泳中得到体现。在彗星试验中，与对照组相比，TSE显著（P<0.001）增加了DNA质量的长宽比。 annexin-V与癌细胞结合的流式细胞术分析进一步支持了TSE的诱导凋亡活性。在培养中研究了TSE对正常人外周血单个核细胞活力和细胞毒性的影响，发现其细胞毒性低于对U937和K562细胞的毒性。本研究结果表明，TSE可能是一种对U937和K562髓系白血病细胞具有抗增殖、细胞毒性和诱导凋亡活性的强效抗肿瘤药物。
