通过MTT分析、光学显微镜观察、Hoechst33342染色质荧光染色、DNA琼脂糖凝胶电泳、罗丹明123染色及钙离子激光扫描显微镜检测等手段,分析了虾夷扇贝毒素(yessotoxin,YTX)对Hela细胞的毒性作用.通过实验发现,正常Hela细胞与不同浓度YTX共孵育24 h后,一定浓度的YTX能对Hela细胞产生毒性作用,使其发生明显凋亡特征,如形态变化、染色质凝缩、DNA片段化以及线粒体膜电位降低.此外,YTX还能造成Hela细胞内胞质钙离子升高.这种毒性作用可能是YTX通过对胞内钙离子浓度的影响进而诱导发生的.%The marine toxin yessotoxin( YTX)and its analogues have been detected in shellfish samples in Chinese coastal areas and they pose a potential threat to public health. Many YTXs toxicity studies have been carried out worldwide, but the process is still indeterminate. In order to study the toxicity of yessotoxin on Hela cells and analyze the possible action mechanism, MTT assay, optical microscope observation, Hoechst 33342 chromatin staining, DNA gel electrophoresis, rhodamine 123 staining, and calcium-sensitive laser scanning confocal microscopy were conducted in this paper. The results demonstrated that YTX could induce a significant reduction of Hela cell viability and the cytotoxicity of the toxin was dose-dependent. Meanwhile, the usual hallmarks of apoptosis could be detected, including chromatin condensation, DNA laddering,and loss of mitochondrial membrane potential. Furthermore, YTX caused cytosolic calcium levels to increase in Hela cells. The change of intracellular calcium concentration may be the main cause of cell apoptosis induced by YTX.
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