姜黄素对人肝癌HepG2细胞毒性的分析

Analysis of Curcumin Toxicity on Human Hepatoma HepG2 Cells

该文结合倒置显微镜、原子力显微镜（AFM）、CCK-8和流式细胞术定性定量研究了姜黄素对人肝癌细胞株HepG2细胞的毒性。AFM探测结果表明，姜黄素能够引起细胞发生不同程度的形变。细胞体积和高度均随细胞形变程度的加深而下降。细胞表面平均粗糙度（Ra）、均方根粗糙度（Rq）和粒径分布均随细胞形变程度的加深而增大。而AFM探测细胞机械性能发现，40μmol／L姜黄素与HepG2细胞共孵育24h后，细胞表面相对粘附力从（708．4±255．7）pN降至（299．84-39．6）pN。CCK-8实验表明，细胞存活率与姜黄素存在时间依赖和剂量依赖关系。20～80μmol／L姜黄素作用细胞24h后，细胞存活率从（72．2±3．8）％降至（10．6±2．3）％；作用48h后，细胞存活率从（53．6±2．7）％降至（8．7±1．8）％。流式细胞术对细胞周期的检测表明，姜黄素能够阻滞肝癌HepG2细胞于G2／M期。该实验对药物引起癌细胞早期凋亡的可视化诊断和研究药物与细胞相互作用有重要意义。

Inverted microscope, atomic force microscope （AFM）, CCK-8 and flow cytometer were used to study the toxicity of curcumin on human hepatoma HepG2 cells qualitatively and quantitatively. The AFM results indicated that curcumin could induce the deformation change of HepG2 cells to varying degrees. The volume and height of the cells decreased with the increase of cell deformation, whereas, the mean roughness（ Ra）, mean-square-root roughness （Rq） and granularity distribution of HepG2 cell surface increased with the increase of cell deformation. Through detecting the mechanical property of the cells by AFM, it was found that the relative adhesion force of the cell surface decreased from （708.4 ± 255.7 ） pN to （299.8 ± 39.6） pN when 40 μmol/L curcumin was incubated with HepG2 cells for 24 h. By means of CCK-8 assay, it was found that the HepG2 cell viability was related with the reaction time and drug concentration. When 20 -80 μmol/L curcumin were incubated with HepG2 cells for 24 h and 48 h, respectively, the cell viability declined from （ 72.2 ± 3.8）% to （10.6±2.3）% and （53.6±2.7）% to （8.7 ±1.8）% , respectively. In addition, the result of cell cycle analysis by flow cytometer showed that curcumin could block the HepG2 cell in G2/M phase. The results obtained above are of significance to the visual diagnosis of drug-induced early apoptosis in cancer cells, and to the study of the drugs - cells interaction.