臭牡丹含药血清对人肝癌MHCC97-H细胞周期和凋亡的影响

Effects of Drug-containing Serum of Clerodendrum Bungei on Cell Cycle and Apoptosis of Human Hepatocellular Carcinoma MHCC97-H

目的观察臭牡丹含药血清对B淋巴细胞瘤/白血病-2(Bcl-2)和Bcl-2相关x蛋白(Bax)蛋白表达及人肝癌MHCC97-H细胞周期、凋亡的影响,探讨其抗肝癌的可能机制。方法15%臭牡丹高、中、低剂量含药血清作用于MHCC97-H细胞,显微镜下观察细胞形态的变化;AnnexinV-FITC/PI双染法检测细胞凋亡;流式细胞仪检测细胞周期;Western blot法检测Bcl-2和Bax蛋白表达的变化。结果不同剂量臭牡丹含药血清作用于细胞后,细胞形态发生稀疏杂乱、折光性增强等改变;流式细胞术结果显示,随含药血清浓度的增加,细胞的凋亡比例和G2/M期细胞比例亦相应增加,S期细胞比例降低,中、高剂量组与模型组比较,差异显著(P<0.05);Western blot法结果显示不同浓度臭牡丹含药血清干预下各组Bax蛋白的表达水平上调,Bcl-2蛋白的表达降低(P<0.01)。结论臭牡丹含药血清可以促进MHCC97-H肝癌细胞凋亡并阻止其细胞周期,机制可能与上调Bax蛋白,下调Bcl-2蛋白,影响Bax/Bcl-2比值有关。

Objective To observe the effects of Clerodendron Bungei drug-containing serum on B lymphocyte/leukemia-2(Bcl-2)and Bcl-2 related X protein(Bax)protein expression and cell cycle and apoptosis of human hepatocarcinoma MHCC97-H,and to explore its possible mechanism.Methods The medicinal serum of 15%Clerodendron Bungei high,medium and low dose drug-containing serum was applied to MHCC97-H cells.The changes in cell morphology were observed using a microscope;The apoptosis was detected by Annexin V-FITC/PI double staining method.The flow cytometry was used to detect cell cycle;Western blot was used to detect Bcl-2 and Bax protein expression.Results After different doses of Clerodendron Bungei drug-containing serum were acting on cells,and the cell morphology was sparse and disordered,and the refractive index was enhanced.The results of flow cytometry showed that with the increase of serum concentration,the proportion of cells in apoptosis and the proportion of cells in G2/M phase increased,and the proportion of cells in S phase decreased.The difference between medium and high dose groups was significantly different from that of the model group(P<0.05).The results of Western blot showed that the expression levels of Bax protein in each group were up-regulated and the expression of Bcl-2 protein was decreased(P<0.01).Conclusion The drug-containing serum of Clerodendron Bungei can promote the apoptosis of MHCC97-H hepatoma cells and prevent its cell cycle.