Wang Fang, Ma Rundi, Yu Lijian
Guangdong Provincial Key Laboratory of Marine Materia Medica, Zhanjiang Ocean University, Guangdong, People's Republic of China.
Cancer Chemother Pharmacol. 2006 Feb;57(3):389-99. doi: 10.1007/s00280-005-0047-y. Epub 2005 Sep 20.
Tubeimoside I (TBMS1), a triterpenoid saponin, isolated from the tubers of Bolbostemma paniculatum, showed potent antitumor and antitumor-promoting effects. The objective of this study is to investigate the role of mitochondria and mitochondria cytochrome c in TBMS1-mediated apoptosis of human cervical carcinoma HeLa cell line.
Viability of HeLa cells was measured by MTT assay. Apoptotic induction by TBMS1 was determined by fluorescence microscopy, flow cytometry and gel electrophoresis of fragmented DNA. Mitochondrial transmembrane potential (Deltapsim) was assayed by flow cytometry. Cytochrome c (Cyt c) was detected by Western blotting.
The results showed that Cyclosporin A (CsA) partly protected HeLa cells from growth inhibitory effect of TBMS1, and partly countered the ability of TBMS1 to rapidly induce apoptosis in HeLa cells, and that TBMS1 decreased Deltapsim and induced Cyt c release by a mechanism inhibited by CsA, and that TBMS1 induced apoptosis of HeLa cells dose-dependently in accordance with increase of cytosolic Cyt c.
TBMS1 opens the permeability transition (PT) pore, thereby decreasing Deltapsim, releasing Cyt c from mitochondria, and further causing a series of events consistent with established mechanistic models of apoptosis.
土贝母苷甲(TBMS1)是一种从土贝母块茎中分离得到的三萜皂苷,具有强大的抗肿瘤及抗肿瘤促进作用。本研究的目的是探讨线粒体及线粒体细胞色素c在TBMS1介导的人宫颈癌HeLa细胞系凋亡中的作用。
采用MTT法检测HeLa细胞的活力。通过荧光显微镜、流式细胞术及DNA片段化凝胶电泳检测TBMS1诱导的凋亡。采用流式细胞术检测线粒体跨膜电位(Δψm)。通过蛋白质免疫印迹法检测细胞色素c(Cyt c)。
结果显示,环孢素A(CsA)部分保护HeLa细胞免受TBMS1的生长抑制作用,部分对抗TBMS1快速诱导HeLa细胞凋亡的能力,且TBMS1降低Δψm并通过一种被CsA抑制的机制诱导Cyt c释放,并且TBMS1根据胞质Cyt c的增加剂量依赖性地诱导HeLa细胞凋亡。
TBMS1打开通透性转换(PT)孔,从而降低Δψm,使Cyt c从线粒体释放,并进一步引发一系列与既定凋亡机制模型一致的事件。