Chen Yun-Jie, Jiang Hai-Tao, Wang Tian-Fei
Department of General Surgery, Ningbo NO. 2 Hospital, Ningbo, China.
Department of General Surgery, Ningbo NO. 2 Hospital, Ningbo, China
Ann Clin Lab Sci. 2019 Jan;49(1):72-78.
This study aims to investigate the effect of different concentrations of docosahexaenoic acid (DHA) on proliferation and apoptosis in HepG2 cell lines, and to research the possible molecular mechanisms.
DHA concentration was 0 g/mL in the negative control group, and 15, 30, 45, 60 and 75 ug/mL, respectively, in the experimental groups. CCK-8 and flow cytometry methods were used to observe the growth inhibition and apoptosis rates of HepG2 cells cultured , which were treated with different concentrations of DHA. The level of β-catenin and c-myc mRNA and protein were measured by real-time PCR and western blot, respectively.
In the concentration range of 0-45 ug/mL, the action time was 24 hours. DHA could inhibit the growth of HepG2 cells, and there were significant differences between the experimental and control groups (<0.01). The same was observed in each of the two groups in experimental groups. As drug concentration or action time increased, results revealed no statistical differences. Furthermore, flow cytometric analysis indicated that DHA could promote HepG2 cell apoptosis; and the apoptosis rate was greatly different between the experimental and control groups (<0.01). The same was observed in each of the two groups in the experimental groups. Real-time PCR could detect low c-myc expression in HepG2 cells disposed by DHA, and c-myc expression was significantly different between the experimental and control groups (<0.01). The same was observed in each of the two groups in the experimental groups. There was no obvious difference in β-catenin expression between the experimental and control groups, and the experimental groups were all identical. Western blot demonstrated that DHA could decrease β-catenin and c-myc protein expression in HepG2 cells.
DHA could promote apoptosis and inhibit the proliferation of HepG2 cells. The possible mechanism was related with the down-regulated protein expression of β-catenin and the mRNA expression of c-myc.
本研究旨在探讨不同浓度的二十二碳六烯酸（DHA）对HepG2细胞系增殖和凋亡的影响，并研究其可能的分子机制。
阴性对照组DHA浓度为0 g/mL，实验组分别为15、30、45、60和75 μg/mL。采用CCK-8法和流式细胞术观察不同浓度DHA处理的HepG2细胞的生长抑制率和凋亡率。分别通过实时PCR和蛋白质印迹法检测β-连环蛋白和c-myc mRNA及蛋白水平。
在0-45 μg/mL浓度范围内，作用时间为24小时。DHA可抑制HepG2细胞生长，实验组与对照组之间存在显著差异（<0.01）。实验组的两组中均观察到同样情况。随着药物浓度或作用时间增加，结果显示无统计学差异。此外，流式细胞术分析表明DHA可促进HepG2细胞凋亡；实验组与对照组之间的凋亡率差异极大（<0.01）。实验组的两组中均观察到同样情况。实时PCR可检测到DHA处理的HepG2细胞中c-myc表达较低，实验组与对照组之间c-myc表达存在显著差异（<0.01）。实验组的两组中均观察到同样情况。实验组与对照组之间β-连环蛋白表达无明显差异，且实验组均相同。蛋白质印迹法表明DHA可降低HepG2细胞中β-连环蛋白和c-myc蛋白表达。
DHA可促进HepG2细胞凋亡并抑制其增殖。可能的机制与β-连环蛋白蛋白表达下调及c-myc mRNA表达有关。
