槲皮素联合顺铂对人卵巢癌SKOV-3细胞增殖及凋亡的影响

Quercetin combined with cisplatin on proliferation and apoptosis in human ovarian cancer SKOV-3 cells

目的探讨槲皮素单独及联合顺铂对人卵巢癌SKOV-3细胞增殖与凋亡的影响,并初步探讨其作用机制。方法选取对数生长期的卵巢癌SKOV-3细胞作为研究对象,实验分6组:阴性对照组(生理盐水),槲皮素低(25μmol/L)、中(50μmol/L)、高剂量组(100μmol/L),顺铂组(10μmol/L),联合组(槲皮素50μmol/L+顺铂10μmol/L)。采用MTT法检测不同浓度槲皮素单独及联合顺铂对SKOV-3细胞的增殖抑制作用;流式细胞仪检测细胞周期分布及细胞凋亡率;免疫细胞化学法检测槲皮素作用于SKOV-3细胞Bcl-2及MtP53蛋白表达水平的变化。结果与阴性对照组相比,槲皮素可明显抑制SKOV-3细胞的增殖,并具有显著的时间和剂量依赖关系,槲皮素高剂量组在72 h的抑制率可达63.49%(P<0.05);流式细胞仪分析发现槲皮素可使G0/G1期细胞明显增多,S期细胞明显减少(P<0.05),提示其将细胞周期阻滞于G1期;槲皮素还可明显诱导SKOV-3细胞凋亡,高剂量组的凋亡率可达29.51%;槲皮素作用后,Bcl-2及MtP53表达则明显降低(P<0.05)。结论槲皮素在体外可抑制SKOV-3细胞增殖,阻滞细胞周期,诱导细胞凋亡,其机制可能与降低凋亡相关蛋白MtP53和Bcl-2的蛋白表达水平有关;槲皮素与顺铂联用有协同作用。

Objective To determine the effect of quercetin combined with cisplatin on the proliferation and apoptosis in SKOV-3 ovarian cancer cells. Methods Ovarian cancer SKOV-3 cells at the logarithmic growth phase were treated with quercetin at 25,50, or 100 μmol/L respectively, with 10 μmol/L DDP, or with 50 μmol/L quercetin and 10 μmol/L DDP together, respectively for 24, 48 and 72 h. The cell proliferation was evaluated by MTT assay, the rate of cell apoptosis and the cell-cycle distribution was detected by flow cytometry, and the expression of P53 and Bcl-2 were analyzed by immunocytochemical assay. Results Quercetin resulted in an decreased of the proliferation in SKOV-3 cells in a dose- and time-dependent manner, and the inhibitory rate in the 100 Ixmol/L quercetin for 72 h was 63.49% （P 〈0.05 ）. Flow cytometry showed that the propor- tions of G0/G1 phase and of S phase were reduced after quercetin exposure （ P 〈 0. 05 ）. Quercetin also increased the apoptotic rate of SKOV-3 cells significantly and the highest apoptotic rate was 29.51% （P 〈 0.05）. Immunocytochemical assay showed that the expression of MtP53 and Bcl-2 proteins in the treated cells was significantly decreased compared with the control （ P 〈 0. 05 ）. Conclusion Quercetin, especially combined with cisplatin, significantly inhibits the proliferation and induces apoptosis of cervical cancer cells, which might be due to the down-regulation of Bcl-2 and MtP53.