米糠油不饱和脂肪酸对肝癌细胞HepG_2克隆及迁移能力的影响

Effect of Unsaturated Fatty Acids from Rice Bran Oil on Cloning and Migration Ability of HepG_2 Cells

本研究采用软琼脂克隆形成和Transwell小室迁移实验观察分析了米糠油不饱和脂肪酸(RBO-UFA)在体外对HepG_2的影响,并通过Western blot检测了凋亡相关因子NF-κB在用RBO-UFA处理前后其表达的变化。Heochst染色结果显示HepG_2经RBO-UFA处理后细胞数量明显减少,胞质浓缩,细胞核内染色质凝集;软琼脂克隆形成检测结果显示HepG_2经RBO-UFA处理后克隆团明显减少,甚至无克隆团出现;迁移检测结果显示亚麻酸和亚油酸抑制HepG_2细胞迁移能力的抑制率分别为47.45±4.60%、41.61±4.08%,其次是米糠油32.85±3.24%,油酸25.55±2.56%(P<0.05)。Western Blot结果显示,RBO-UFA处理后HepG_2细胞全蛋白中NF-κB表达水平显著低于正常对照组,米糠油、油酸、亚油酸、亚麻酸处理组与正常对照组相比,其相对表达量分别降低了0.46±0.017、0.058±0.018、0.25±0.005、0.47±0.011;而在核蛋白中NF-κB表达水平均高于正常对照组,相对表达量分别增加了0.59±0.036、0.053±0.040、0.28±0.043、0.59±0.007(P<0.05)。结果表明米糠油不饱和脂肪酸(RBO-UFA)对肝癌细胞HepG_2的克隆形成及迁移具有一定的抑制作用。

The in vitro effects of unsaturated fatty acids of rice bran oil (RBO-UFA) on HepG2cells were examined by soft agar colony formation and transwell migration assays. Changes in the expression of the apoptosis-related factor, NF-κB, before and after RBO-UFA treatment were analyzed by western blot assay. Hoechst staining indicated that the number of HepG2cells after RBO-UFA treatment was significantly decreased and the cytoplasm and nuclear chromatin were condensed. The soft agar colony formation assay showed that the HepG2colonies were markedly reduced after RBO-UFA treatment and no colonies appeared. The migration assay indicated that the inhibition rates of linolenic acid and linoleic acid on the migration ability of HepG2cells were up to 47.45 ± 4.60% and 41.61±4.08%, respectively, followed by that of rice bran oil (32.85 ± 3.24%) and that of oleic acid (25.55 ± 2.56%) (P 2hepatic cancer cells. © 2015, Editorial Board of Modern Food Science and Technology. All right reserved.