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HepG2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HepG2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HepG2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HepG2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HepG2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HepG2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 HepG2 cells 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 HepG2 cells after RBO-UFA treatment was significantly decreased and the cytoplasm and nuclear chromatin were condensed. The soft agar colony formation assay showed that the HepG2 colonies 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 HepG2 cells 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 < 0.05). The western blot results showed that NF-κB expression levels in total proteins from RBO-UFA treated groups were significantly lower than that of the normal control group. Compared with the control group, the relative expression levels of NF-κB in HepG2 cells treated with rice bran oil, oleic acid, linoleic acid, and linolenic acid were reduced by 0.46 ± 0.017, 0.058 ± 0.018, 0.25 ± 0.005, and 0.47 ± 0.011, respectively. On the other hand, compared with the control group, NF-κB expression in the RBO-UFA treated groups in the nuclear protein fraction was higher than that in the normal control group. The relative expression levels of NF-κB in HepG2 cells treated with rice bran oil, oleic acid, linoleic acid and linolenic acid were increased by 0.59 ± 0.036, 0.053 ± 0.040, 0.28 ± 0.043, and 0.59 ± 0.007 (P < 0.05), respectively. In summary, RBO-UPA treatment presented an inhibitory effect on the migration and clonogenicity of HepG2 hepatic cancer cells.

国家自然科学基金（31201348）；湖南省自然科学基金（13JJ4086）；湖南省农业成果转化项目（2013NK4002）；长沙市科技计划项目（K1403039-21）