臭牡丹含药血清抑制人肝癌MHCC97-H细胞侵袭与迁移的作用及其机制研究

Inhibitory effect and mechanism of Clerodendrum bungei-containing serum on invasion and migration of human hepato cellular carcinoma cell line MHCC97-H

目的探讨臭牡丹含药血清对人肝癌细胞(MHCC97-H细胞)迁移与侵袭能力的影响及其机制。方法按照体重将SD大鼠随机分成4组:模型组和低、中、高3个剂量实验组,每组5只。低、中、高3个剂量实验组给予臭牡丹提取物0.55,1.09,2.18 g·kg^-1;模型组给予0.9%Na Cl,每日灌胃2次,连续3 d。腹主动脉取血后分离血清,制备各组含药血清,体外培养MHCC97-H细胞。Transwell实验检测细胞的迁移能力;免疫印迹法检测磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路中的磷酸Akt(p-Akt)和PI3K相关蛋白表达的影响。结果模型组和低、中、高3个剂量实验组的迁移细胞数分别为(256.0±28.6),(181.0±15.7),(118.0±14.7)和(66.0±12.4)个,3个实验组与模型组相比,差异均有统计学意义(均P<0.01)。模型组和低、中、高3个剂量实验组的PI3K蛋白相对表达量(灰度值)分别为0.68±0.03,0.65±0.02,0.48±0.01和0.15±0.01;上述这4组的p-Akt蛋白相对表达量(灰度值)分别为0.37±0.04,0.33±0.02,0.25±0.01和0.09±0.01,中、高2个剂量实验组与模型组相比,差异均有统计学意义(均P<0.01)。结论臭牡丹含药血清具有抑制肝癌MHCC97-H细胞迁移和侵袭的作用,其作用机制可能与PI3K/Akt信号通路相关。

Objective To investigate the effects of serum containing Clerodendrum bungei on the migration and invasion of human hepatoma cellular carcinoma MHCC97-H cells,and to study its preliminary mechanism.Methods SD rats were randomly divided into 4 groups according to body weight:model group and experimental-L,experimental-M,experimental-H groups,each group had 5 rats.The experimental-L,experimental-M,experimental-H groups were given 0.55,1.09,2.18 g·kg^-1 of the extract of Clerodendrum bungei;model group was given normal saline;twice a day for 3 days.The serum was separated after blood was taken from the abdominal aorta.Each group of drug-containing serum was prepared,and MHCC97-H cells were cultured in vitro.The cell invasion and migration ability were detected by Transwell.The Phosphoprotein kinase B(p-Akt)and Phosphatidylinositol 3-kinase(PI3 K)-related protein expressionf PI3 K/Akt signaling pathway were detected by Western blot.Results The number of migration transmembrane cells in model group and experimental-L,experimental-M,experimental-H groups were(256.0±28.6),(181.0±15.7),(118.0±14.7),(66.0±12.4),respectively,comparison between three doses experimental groups and model group,the difference was significantly(all P<0.05).The relative expression levels of PI3 K protein(grey value)in model group and experimental-L,experimental-M,experimental-H groups were 0.68±0.03,0.65±0.02,0.48±0.01,0.15±0.01;the relative expression levels of p-Akt protein(grey value)in the above 4 groups were 0.37±0.04,0.33±0.02,0.25±0.01,0.09±0.01;comparison between experimental-M,experimental-H groups and model group,the difference of the factors were significantly(all P<0.01).Conclusion The medicinal serum of Clerodendron bungei can inhibit the migration and invasion of liver cancer MHCC97-H cells,and its mechanism may be related to PI3 K/Akt signaling pathway.