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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。
In this study, soft agar colony formation and Transwell chamber migration experiments were used to analyze the effect of RBO-UFA on HepG_2 in vitro, and the apoptosis-related factor NF-κB was detected by Western blot on RBO-UFA. Changes in expression before and after treatment. Heochst staining results showed that HepG_2 cells treated with RBO-UFA significantly reduced the number of cells, condensed the cytoplasm, and aggregated the chromatin in the nucleus. The results of soft agar colony formation showed that the HepG_2 treated with RBO-UFA significantly reduced the number of cloned clumps, and even no clones appeared. The results of migration assay showed that the inhibitory rates of linolenic acid and linoleic acid inhibited the migration ability of HepG 2 cells were 47.45±4.60% and 41.61±4.08%, followed by rice bran oil 32.85±3.24% and oleic acid 25.55±2.56% (P<0.05). ). Western Blot results showed that the expression of NF-κB in HepG_2 cells after RBO-UFA treatment was significantly lower than that in the normal control group. The relatives of rice bran oil, oleic acid, linoleic acid, and linolenic acid were compared with the normal control group. The expression levels were decreased by 0.46±0.017, 0.058±0.018, 0.25±0.005, and 0.47±0.011, respectively. The expression of NF-κB in the nuclear protein was higher than that in the normal control group. The relative expression levels were increased by 0.59±0.036 and 0.053±, respectively. 0.040, 0.28±0.043, 0.59±0.007 (P<0.05). The results showed that RBO-UFA had a certain inhibitory effect on the colony formation and migration of HepG_2 cells.