Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China
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 highdose 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.
TIAN Lu, CHEN Ying, ZHANG Yang.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[J].中国比较医学杂志,2023,33(7):78~84.复制