Department of Gastroenterology, Qingdao Hiser Medical Group, Qingdao, China.
Eur Rev Med Pharmacol Sci. 2018 Jul;22(14):4500-4508. doi: 10.26355/eurrev_201807_15504.
Omega-3 polyunsaturated fatty acid (ω-3 PUFA) has been found to possess anti-cancer potential in previous studies. However, the underlying mechanism of ω-3 PUFA in protecting hepatocarcinoma has not been fully elucidated. This study aims to explore the function of ω-3 PUFA in the development of hepatocarcinoma and its potential mechanism.
In this study, human hepatocarcinoma cell line Hep G2 was treated with ω-3 PUFA. Cell counting kit-8 (CCK-8) and cell cloning assay were applied to detect the proliferation of Hep G2 cells. In addition, flow cytometry was performed to analyze the cell cycle and apoptosis rate. At the same time, the effect of ω-3 PUFA on invasion and metastasis of hepatocarcinoma cells were analyzed by transwell assay. Moreover, protein levels of key factors in Wnt/β-catenin pathway were detected by Western blot.
Cell proliferation of Hep G2 cells was decreased after ω-3 PUFA treatment in a time- and dose-dependent manner. CCK-8 assay showed that the IC50 value was 12.8 ± 0.67 μmol/L, 8.8 ± 0.43 μmol/L and 4.6 ± 0.42 μmol/L after ω-3 PUFA treatment for 24 h, 48 h and 72 h, respectively. Besides, ratio of Hep G2 cells blocked at G2/M phase after ω-3 PUFA treatment (5 μmol/L, 10 μmol/L and 20 μmol/L) was increased in a dose-dependent manner (p<0.05). Meanwhile, ω-3 PUFA could increase cell apoptosis (p<0.05) and inhibit cell proliferation. In addition, ω-3 PUFA reduced protein expressions of total, cytoplasmic and nuclear β-catenin in Hep G2 cells, indicating that the Wnt/β-catenin pathway is inhibited. Decreased expression levels of Dvl-2, Dvl-3, GSK-3β (p-ser9), c-myc and survivin, and increased expression levels of GSK-3 (p-tyr216) and Axin-2 were observed in Hep G2 cells treated with ω-3 PUFA, but no significant alteration in total GSK-3β protein level was observed (p>0.05).
Omega-3 PUFA regulates the malignant progression of hepatocarcinoma by inhibiting proliferation and promoting apoptosis of hepatocarcinoma cells via Wnt/β-catenin signaling pathway.
已有研究发现，ω-3 多不饱和脂肪酸（ω-3PUFA）具有抗癌潜力。然而，ω-3PUFA 保护肝癌的潜在机制尚未完全阐明。本研究旨在探讨 ω-3PUFA 在肝癌发展中的作用及其潜在机制。
本研究用人肝癌细胞系 Hep G2 处理 ω-3PUFA。应用细胞计数试剂盒-8（CCK-8）和细胞克隆实验检测 Hep G2 细胞的增殖。此外，通过流式细胞术分析细胞周期和细胞凋亡率。同时，通过 Transwell 实验分析 ω-3PUFA 对肝癌细胞侵袭和转移的影响。此外，通过 Western blot 检测 Wnt/β-catenin 通路中关键因子的蛋白水平。
随着时间和剂量的增加，ω-3PUFA 处理后 Hep G2 细胞的增殖减少。CCK-8 检测结果显示，ω-3PUFA 处理 24 h、48 h 和 72 h 后 Hep G2 细胞的 IC50 值分别为 12.8±0.67 μmol/L、8.8±0.43 μmol/L 和 4.6±0.42 μmol/L。此外，随着 ω-3PUFA 浓度的增加，Hep G2 细胞被阻滞在 G2/M 期的比例呈剂量依赖性增加（p<0.05）。同时，ω-3PUFA 能促进细胞凋亡（p<0.05）并抑制细胞增殖。此外，ω-3PUFA 降低了 Hep G2 细胞中总、细胞质和核 β-catenin 的蛋白表达，表明 Wnt/β-catenin 通路被抑制。在经 ω-3PUFA 处理的 Hep G2 细胞中，观察到 Dvl-2、Dvl-3、GSK-3β（p-ser9）、c-myc 和 survivin 的表达水平降低，而 GSK-3（p-tyr216）和 Axin-2 的表达水平升高，但总 GSK-3β 蛋白水平无明显变化（p>0.05）。
ω-3PUFA 通过抑制 Wnt/β-catenin 信号通路，调节肝癌细胞的增殖和促进其凋亡，从而调节肝癌的恶性进展。
