冬凌草甲素抑制体内外卵巢癌生长的作用

In Vitro and in Vivo Anti-Tumor Effect of Oridonin on Ovarian Cancer

目的:探讨冬凌草甲素对体内外卵巢癌生长的影响及其作用机制。方法:冬凌草甲素作用人卵巢癌细胞株HO-8910PM后,MTT法检测细胞增殖;流式细胞术检测细胞凋亡;Westernblot检测卵巢癌细胞中核因子(NF)-κB和X连锁凋亡抑制蛋白(XIAP)表达;建立起裸鼠卵巢癌皮下移植瘤模型,观察冬凌草甲素对裸鼠卵巢癌皮下移植瘤生长的影响;免疫组织化学法检测肿瘤组织中Ki-67、NF-κB和XIAP的阳性表达。结果:HO-8910PM细胞经不同浓度冬凌草甲素(10、20、40μmol·L-1)作用24h后,细胞存活率分别为(80.14±9.84)%、(71.68±6.51)%和(64.58±5.24)%,均低于对照组(96.12±4.23)%,差异有统计学意义(P<0.05)。冬凌草甲素(40μmol·L-1)作用HO-8910PM细胞24h后,诱导(15.9±3.4)%的HO-8910PM细胞发生早期凋亡,高于对照组的(1.7±0.3)%,差异有统计学意义。3种浓度(10、20、40μmol·L-1)的冬凌草甲素作用24h,均可抑制HO-8910PM细胞中NF-κB的表达;而低浓度(10μmol·L-1)冬凌草甲素对HO-8910PM细胞中XIAP蛋白无明显抑制作用,高浓度冬凌草甲素(20和40μmol·L-1)明显抑制XIAP蛋白的表达。冬凌草甲素可显著抑制裸鼠卵巢癌皮下移植瘤生长,实验组中Ki-67、NF-κB和XIAP的表达强度均低于对照组(P<0.01)。结论:冬凌草甲素可显著抑制体内外卵巢癌的生长,该作用机制可能是冬凌草甲素通过抑制NF-κB及其调控蛋白XIAP的表达来抑制卵巢癌的生长。

Objective：To investigate in Vitro and in Vivo effects and the mechanism of oridonin on the growth of ovarian cancer . Methods：After human ovarian cancer cell line HO-8910PM was treated with different concentrations of oridonin, the cellular proliferation was detected by MTT assay. The flow cytometry was used to determine the apoptosis in ovarian cancer cells. Western blot was used to detect expressions of nuclear factor （NF）-κB and x-linked？inhibitor of apoptosis protein？（XIAP） in ovarian cancer cells. Furthermore, HO-8910PM cells were injected subcutaneously into nude mice to establish xenograft model. The tumor weight and inhibition rate were evaluated respectively after treatment with oridonin in nude mice. The positive expressions of Ki-67, NF-κB and XIAP in tumor tissues were detected by immunohistochemistry method. Results： The proliferation of ovarian cancer cells was inhibited significantly by oridonin. The cell viability values were （80.14±9.84）%, （71.68±6.51）% and （64.58±5.24）% respectively after treatment with the concentrations of oridonin 10, 20 and 40 μmol/L for 24 h, which were significantly lower than those （96.12±4.23）% in control group （P 〈 0.05）. Treatment with oridonin （40 μmol·L-1） for 24 h induced an early apoptosis with （15.9±3.4）% in HO-8910PM cells, which was significantly higher than that of control （1.7±0.3）%. Treatment with oridonin （10, 20 and 40 μmol·L-1） for 24 h in HO-8910PM cells induced the expression of NF-κB protein. The lower concentration of oridonin （10 μmol·L-1） had no inhibitory effect on the expression of XIAP in HO-8910PM cells. Meanwhile, the higher concentration of oridonin （20 and 40 μmol·L-1） significantly inhibited the expression of XIAP. Furthermore, the subcutaneous？tumor growth was significantly inhibited by oridonin in nude mice. The positive expressions of Ki-67, NF-κB and XIAP were significantly decreased in experimental group than those of control group （P 〈 0.01）. Conclusion： Oridonin exerts in vitro and in vivo anti-tumor activity in ovarian cancer , which may be related to the down-regulated levels of NF-κB and XIAP.