In vitro ultramorphological assessment of apoptosis induced by zerumbone on (HeLa).
In vitro ultramorphological assessment of apoptosis induced by zerumbone on (HeLa).
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DOI：
10.1155/2009/769568
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发表时间：
2009
影响因子：
--
通讯作者：
Mohan S
中科院分区：
文献类型：
--
作者：
Abdel Wahab SI;Abdul AB;Alzubairi AS;Mohamed Elhassan M;Mohan S
Zerumbone(Zerumbone)，一种潜在的抗癌化合物，从生姜的新鲜根茎中分离得到。采用四甲基偶氮唑盐比色法测定了ZER对人宫颈癌细胞(HeLa)、乳腺癌细胞、卵巢癌细胞和中国仓鼠卵巢正常细胞的细胞毒作用。采用荧光显微镜(AO/PI双重染色)、扫描和透射电子显微镜(SEM和TEM)以及细胞凋亡促进酶caspase-3的比色分析，研究了ZER对HeLa细胞的促凋亡作用。四甲基偶氮唑盐比色法结果显示，ZER对正常细胞的作用弱于对癌细胞的作用。ZER对HeLa细胞的IC50最低。细胞学观察显示核固缩、染色质固缩、细胞皱缩、多核、线粒体脊异常、膜起泡、空洞、胞质突起和凋亡小体形成，这一点经AO/PI、扫描电子显微镜和透射电子显微镜双重染色共同证实。荧光显微镜下200个细胞差异计数的统计分析(双尾t检验)显示，经处理的HeLa细胞与未处理的HeLa细胞的凋亡数有显著差异。此外，ZER还增加了经处理的HeLa细胞上caspase-3的细胞水平。可以认为，ZER能够产生与细胞凋亡相对应的细胞死亡的独特形态特征。
Zerumbone (ZER), a potential anticancer compound, isolated from the fresh rhizomes of Zingiber zerumbet. In this investigation, the cytotoxic properties of ZER were evaluated, on cancer cells of human cervix (HeLa), breast and ovary, and normal cells of Chinese Hamster ovary, using MTT assay. Apoptogenic effects of ZER on HeLa were studied using fluorescence microscopy (AO/PI double staining), scanning and transmission electron microscopy (SEM and TEM), and colorimetric assay of the apoptosis promoter enzyme, caspase-3. The results of MTT assay showed that ZER has less effect on normal cells compared to cancer cells. The lowest IC50 of ZER was observed on HeLa cells. Cytological observations showed nuclear and chromatin condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, holes, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by double staining of AO/PI, SEM and TEM. Statistical analysis (two-tailed t-test) of differential counting of 200 cells under fluorescence microscope revealed significant difference in apoptotic cells populations between treated and untreated HeLa cells. In addition, ZER has increased the cellular level of caspase-3 on the treated HeLa cells. It could be concluded that ZER was able to produce distinctive morphological features of cell death that corresponds to apoptosis.