环肽a-amanitin对人肝癌细胞SMMC-7721的体外毒性作用研究

Inhibitory Effect of Cyclopeptide α-amanitin on Human Hepatoma SMMC-7721 Cells

目的研究毒蕈环肽a-amanitin对人肝癌细胞SMMC-7721的体外毒性作用。方法运用MTT法研究α-aman-itin不同浓度和不同作用时间对体外传代培养的人肝癌细胞SMMC-7721的抑制增殖作用,再进行环肽α-amanitin对细胞毒性作用的数学模型拟合。结果研究发现,α-amanitin的使用剂量与细胞存活率存在负相关。7.12×10-4mmol/L和7.12×10-10mmol/Lα-amanitin对SMMC-7721作用24 h后,细胞存活率分别为71.25%和87.47%,作用效果分别与10 mmol/L和2.5 mmol/L环磷酰胺相似。SMMC-7721细胞存活率平方根的反正弦值与α-amanitin浓度的对数变换值的相关和回归分析结果显示二者显著性相关,曲线拟合结果显示三次曲线模型拟合最好,回归方程为Y=31.257 7-12.902X-1.5117X2-0.0616X3。对SMMC-7721细胞存活率平方根的反正弦值与α-amanitin作用时间进行相关和回归分析,二者显著性相关,三次曲线模型拟合最好,对应回归方程为Y=89.668 5-8.616 6X+1.791X2-0.1271X3。结论环肽a-amanitin对人肝癌细胞SMMC-7721体外增殖有显著抑制作用,且单位摩尔数的α-amanitin抑制作用远强于单位摩尔数的环磷酰胺。

Objective To study the inhibitory effect of cyclopeptide α-amanitin on human hepatoma SMMC-7721 cells.Methods The SMMC-7721 cells were treated with different concentration and duration of α-amanitin.The cell survival rate（CRS） of SMMC-7721 cells was examined by MTT assay in vitro,and the relative results were fitted with mathematical model.Results The cell survival rate was reversely correlated with the concentration of α-amanitin.The cell growth was restrained strikingly when the cells were treated by 7.12×10^-10 mmol/L α-amanitin for 24 h.The cell survival rate were 71.25% and 87.47% after treating with 7.12×10^-4 mmol/L and 7.12×10^-10 mmol/L α-amanitin for 24 h,respectively.The arcsine transformation of the CSR square root and the logarithmic transformation of α-amanitin concentration were correlated and regressed.The curve fitting showed that the cubic model was the best one.The regression equation was Y=31.257 7-12.902X-1.511 7X^2-0.061 6X^3.The arcsine transformation of the CSR square root and the action time of α-amanitin were correlated and regressed.The correlation was also significant and the cubic model was the best one.The cubic regression equation was Y=89.668 5-8.6166X＋1.791X^2-0.1271X^3.Conclusion The cyclopeptide α-amanitin has significant inhibitory effect on human hepatoma SMMC-7721 cells,and its toxicity is much stronger than cyclophosphamide.