姜黄素超分子包合物抑制HepG2肝癌细胞的增殖

Inhibition Effect of Curcumin/β-cyclodextrin Polymer Inclusion Complex on the Proliferation of HepG2 Hepatocellular Carcinoma Cells

对姜黄素超分子包合物抑制HepG2肝癌细胞增殖的作用进行研究。采用MTT法考察不同浓度的姜黄素超分子包合物(40～640μg/mL)处理HepG2细胞不同时间(24h、48h和72h)后对其细胞存活率的影响。然后,采用流式细胞术和测定Caspase 3/8/9酶活来探讨姜黄素超分子包合物抑制HepG2细胞增殖的作用机理。实验结果表明,随着姜黄素超分子包合物处理时间和浓度的增加,HepG2细胞的存活率呈现出逐渐下降的趋势,当处理时间为72h,姜黄素超分子包合物浓度为640μg/mL时,细胞的存活率达到最低为9.81%±1.00%。进一步的流式细胞术分析得知,HepG2细胞内的凋亡细胞数目随着姜黄素超分子包合物浓度的增加而增加,当细胞经640μg/mL姜黄素超分子包合物处理72h后,其细胞内凋亡细胞的数目(Sub-G1)达到最高为93.43%。通过对Caspase 3/8/9的酶活进行检测发现,Caspase 3/8/9的酶活也是随着姜黄素超分子包合物浓度的增加而增加,当640μg/mL姜黄素超分子包合物处理细胞72h后,Caspase 3/8/9的酶活达到最大。进一步地Western blot实验结果也表明,随着姜黄素超分子包合物浓度的增加,Caspase 3/8/9的蛋白表达水平呈升高趋势。上述实验结果表明,姜黄素超分子包合物是通过线粒体途径和死亡受体途径来诱导细胞发生凋亡从而抑制HepG2细胞增殖。

In this paper,the inhibition effect of curcumin/β-cyclodextrin polymer inclusion complex on the proliferation of HepG2 hepatocellular carcinoma cells was studied.MTT assay was used to investigate the effects of curcumin/β-cyclodextrin polymer inclusion complex with the different concentrations(40~640μg/mL)and different periods of times(24 h,48 h,and 72 h)on the cell viability of HepG2 cells.Then,the flow cytometry and measuring Caspase 3/8/9 activity were used to investigate the inhibiting mechanism of curcumin/β-cyclodextrin polymer inclusion complex on the proliferation of HepG2 cells.The results showed that with the increase in the treatment times and concentrations of curcumin/β-cyclodextrin polymer inclusion complex,the cell activity of HepG2 cells was gradually decreased.When the HepG2 cells were exposed to curcumin/β-cyclodextrin polymer inclusion complex(640μg/mL)for 72 h,the cell activity reached the lowest as 9.81%±1.00%.Further flow cytometry analysis revealed that with the increase in the concentration of curcumin/β-cyclodextrin polymer inclusion complex,the number of apoptotic cells in HepG2 cells was gradually increased.When the cells were exposed to inclusion complex(640μg/mL)for 72 h,the number of apoptotic cells(sub-G1)in HepG2 cells reached a maximum of 93.43%.The activity of Caspase 3/8/9 was also increased with the increasing concentration of curcumin/β-cyclodextrin polymer inclusion complex.The activity of caspase3/8/9 reached the maximum after the cells was exposed to inclusion complex(640μg/mL)for 72 h.The results of western blot also showed that the protein expression level of Caspase 3/8/9 increased with the increase of the curcumin/β-cyclodextrin polymer inclusion complex concentration.The above results indicated that curcumin/β-cyclodextrin polymer inclusion complex inhibited HepG2 cells proliferation through mitochondrial pathways and death receptor pathways to induce HepG2 apoptosis.