扶正抗癌汤含药血清对人卵巢癌HO-8910PM细胞增殖、凋亡与周期进程的影响

Effects of serum containing Fuzheng kangai decoction on proliferation, apoptosis and cell cycle progression of human ovarian carcinoma HO-8910PM cells

目的探讨扶正抗癌汤(Fuzheng kangai decoction,FZKAD)含药血清对人卵巢癌HO-8910PM细胞增殖、凋亡与周期进程的影响。方法SD大鼠40只随机分为对照组及FZKAD低(4.725 g/kg)、中(9.45 g/kg)、高(18.9 g/kg)剂量组,灌胃给药7 d后制备含药血清,用于HO-8910PM细胞的培养。分别采用噻唑蓝(MTT)实验检测细胞增殖能力,平板克隆形成实验检测克隆形成能力,膜联蛋白V(Annexin V)-异硫氰酸荧光素/碘化丙啶(FITC/PI)双染法流式细胞术检测细胞凋亡,PI单染法流式细胞术检测细胞周期进程,Western blot法检测蛋白表达水平。结果与对照组比较,除低、中剂量组处理24 h后的细胞存活率无显著性降低外(P>0.05),其余各组处理24、48及72 h后的细胞存活率均显著性降低(P<0.05或P<0.01),且各剂量组细胞克隆形成能力也显著降低(P<0.01);与对照组比较,低、中、高剂量组的细胞凋亡率及P53、Bcl-2相关X蛋白(Bax)表达水平增加(P<0.05或P<0.01),而B淋巴细胞瘤-2(Bcl-2)蛋白表达水平降低(P<0.01);与对照组比较,低、中、高剂量组G0/G1期细胞百分比增加(P<0.05或P<0.01),而S期、G2/M期细胞百分比及细胞周期蛋白D1(Cyclin D1)、周期蛋白依赖性激酶4(CDK4)、CDK6蛋白表达水平降低(P<0.05或P<0.01)。结论FZKAD含药血清对人卵巢癌HO-8910PM细胞的增殖具有抑制作用,该作用与诱导凋亡及周期进程阻滞有关。

Objective To investigate the effects of serum containing Fuzheng Kangai Decoction(FZKAD)on the proliferation,apoptosis,and cell cycle progression in human ovarian carcinoma HO-8910PM cells.Methods Forty Sprague-Dawley rats were divided randomly into control and low-dose(4.725 g/kg),medium-dose(9.45 g/kg),and high-dose(18.9 g/kg)FZKAD groups.After 7 days of intragastric administration,serum containing FZKAD was prepared and used for the culture of HO-8910PM cells.Cell proliferation was detected by MTT assay,colony-formation ability was detected by clone formation assay,apoptosis was detected by flow cytometry using Annexin V-fluorescein isothiocyanate/propidium iodide double-staining,cell cycle progression was detected by flow cytometry using propidium iodide single-staining,and protein expression levels were detected by Western blot.Results There was no significant difference in cell survival rates among the low-dose,medium-dose,and control groups after 24 h(P>0.05).However,cell survival rates in the other groups were significantly decreased after 24,48 and 72 h(P<0.05 or P<0.01),and the colony-formation abilities were also significantly decreased in each dosage group(P<0.01).Compared with the control group,the apoptosis rates and protein expression levels of P53 and Bax were significantly increased in the low-dose,medium-dose,and high-dose groups(P<0.05 or P<0.01),while the protein expression level of Bcl-2 was decreased(P<0.01).The percentage of cells in G0/G1 stage was significantly increased in the low-dose,medium-dose,and high-dose groups compared with the control group(P<0.05 or P<0.01),while the percentages of cells in S and G2/M stages and protein expression levels of cyclin D1 and cyclin-dependent kinases 4 and 6 were decreased(P<0.05 or P<0.01).Conclusions Serum containing FZKAD can inhibit the proliferation of human ovarian cancer HO-8910PM cells,related to the induction of apoptosis and cell cycle arrest.