鳖甲煎丸化裁方及其拆方对肝癌H22细胞的影响

Effects of Modified Biejiajian Pill and Its Separated Prescriptions on Hepatoma H22 Cells of Liver Cancer

目的探讨鳖甲煎丸化裁方及其拆方(软坚散结方、扶正方)对肝癌H22细胞增殖、侵袭、迁移及上皮间质转化(EMT)的影响。方法将50只健康SD大鼠随机分为空白组,鳖甲煎丸化裁方低、中、高浓度组,软坚散结方低、中、高浓度组,扶正方低、中、高浓度组,每组5只,制备含药血清;采用CCK8法检测各组大鼠含药血清干预肝癌H22细胞12 h、24 h、48 h后的增殖影响;采用Transwell检测鳖甲煎丸化裁方及其拆方对肝癌H22细胞迁移与侵袭能力的影响;免疫荧光检测Vimentin与E-cadherin蛋白的表达,评估鳖甲煎丸化裁方及其拆方对H22肝癌细胞EMT进程的影响。结果各给药组干预12 h、24 h后,与空白组比较,鳖甲煎丸化裁方、软坚散结方、扶正方的低、中、高浓度组均可抑制H22肝癌细胞的增殖,差异有统计学意义(P<0.05)。各给药组干预48 h后,与空白组比较,除鳖甲煎丸化裁方低浓度组外,其余各组均可抑制H22肝癌细胞的增殖,差异有统计学意义(P<0.05)。与空白组比较,鳖甲煎丸化裁方高浓度组及其拆方高浓度组均可抑制H22细胞的迁移和侵袭能力,差异有统计学意义(P<0.05),与鳖甲煎丸高浓度组比较,软坚散结高浓度组、扶正高浓度组的细胞迁移和侵袭数相对较高,鳖甲煎丸高浓度组中迁移和侵袭细胞数量最少,表明鳖甲煎丸化裁方高浓度组较其拆方高浓度组抑制肝癌H22细胞的迁移和侵袭能力更明显。与空白组比较,鳖甲煎丸化裁方高浓度组及其拆方高浓度组可提高E-cadherin相对荧光表达量,降低Vimentin的表达,差异有统计学意义(P<0.05)。结论鳖甲煎丸化裁方及其拆方可抑制H22肝癌细胞的增殖、侵袭、迁移及EMT的进程,鳖甲煎丸化裁方较拆方效果更显著。

Objective To investigate the effects of Biejiajian Pills and its decomposed formulas(Ruanjian Sanjie Recipe,Fuzheng Recipe)on the proliferation,invasion,migration and epithelial-mesenchymal transition(EMT)of liver cancer H22 cells.Methods 50 healthy SD rats were randomly divided into blank groups,low,medium,and high dose groups of modified Biejiajian Pills,Ruanjian Sanjie Recipe,and Fuzheng Recipe to prepare drug-containing serum,5 rats in each group.CCK8 method was used to detect the proliferation of hepatoma H22 cells after 12 h,24 h and 48 h intervention with drug-containing serum in each group.Transwell was used to detect the effect of the modified Biejia Jianwan recipe and its separated prescriptions on the migration and invasion of hepatoma H22 cells.The expression of Vimentin and E-cadherin protein was detected by immunofluorescence,and the effect of modified Biejia Jianwan recipe and its separated prescriptions on EMT process of hepatoma H22 cells was evaluated.Results After 12 hours and 24 hours of intervention in each administration group,compared with the blank group,the low,medium and high concentration groups of Biejiajian Pill,Huajia recipe,Ruanjian Sanjie Formula,and Fuzheng Formula could inhibit the proliferation of H22 liver cancer cells,the difference is statistically significant(P<0.05).After 48 h of intervention in each administration group,compared with the blank group,except for the low-concentration group of Biejiajian Pills,the other groups could inhibit the proliferation of H22 liver cancer cells,and the difference was statistically significant(P<0.05).Compared with the blank group,the high-concentration group of Biejiajian Pills and the high-concentration group of its disassembled prescriptions can inhibit the migration and invasion of H22 cells,and the difference is statistically significant(P<0.05),which is higher thanBiejiajian Pills.Compared with the concentration groups,the number of cell migration and invasion was relatively higher in the Ruanjian Sanjie high concentration group and the Fuzheng high concentration group,and the number of migration and invasion cells in the high concentration group of Biejiajian Pill was the least,indicating that Biejiajian Pills had a high concentration.Compared with the disassembled high concentration group,the group inhibited the migration and invasion of liver cancer H22 cells more significantly.Compared with the blank group,the high-concentration group of Biejiajian Pills and its disassembled high-concentration group can increase the relative fluorescence expression of E-cadherin and reduce the expression of Vimentin,and the difference is statistically significant(P<0.05).Conclusion Modified Biejiajian pill and its separated prescriptions can inhibit the proliferation,invasion,migration,and EMT process of hepatoma H22 cells.Modified Biejiajian pill is more significant than its separated prescriptions.