β-羟基异戊酰紫草素联合顺铂对SKOV_3细胞活力的影响

β-HIVS Combined Cisplatin Inhibited Activities of Human Ovarian Cancer Cell Line SKOV_3 in vitro

目的观察β-羟基异戊酰紫草素（β-HIVS）联合顺铂对人卵巢癌细胞株SKOV_3活力的影响,并探讨其可能的发生机制。方法设立空白对照组及6个不同浓度β-HIVS组（2~30μmol/L）,应用四甲基偶氮唑蓝（MTT）比色法测定不同浓度β-HIVS对SKOV_3细胞活力的影响。设立顺铂单用组（10、20、40μmol/L）和选择〈IC_（50）的β-HIVS 3个浓度（0.25、1、2.5μmol/L）协同顺铂组,应用MTT比色法测定β-HIVS协同顺铂对SKOV_3细胞活力的影响。应用Western blot法检测不同浓度β-HIVS作用后BcI-2、Bax蛋白表达。结果不同浓度β-HIVS（2～30μmol/L）均可抑制SKOV_3细胞活力,并呈一定的浓度依赖性,测得半数抑制率IC_（50）为7.37μmol/L,与空白对照组比较,差异均有统计学意义（P〈0.05）。β-HIVS（1、2.5μmol/L）与顺铂联合应用,与顺铂单用组比较,差异亦有统计学意义（P〈0.05,P〈0.01）。协同作用对β-HIVS呈现浓度依赖性。Western bIot检测结果表明：不同浓度β-HIVS（5、7.5、10μmol/L）可明显上调SKOV_3细胞中Bax蛋白表达水平,同时抑制Bcl-2蛋白表达,与对照组比较,差异有统计学意义（P〈0.01）。结论β-HIVS对人卵巢癌细胞株SKOV_3体外活力有明显的抑制作用,且呈浓度依赖性。在一定浓度范围内,β-HIVS对顺铂具有协同增敏作用,且随β-HIVS浓度的增高,这种协同作用表现的更为显著。协同作用可能通过β-HIVS上调Bax蛋白表达和抑制Bcl-2蛋白表达,从而加速诱导SKOV_3凋亡。

Objective To study the effect of β-hydroxyisovaleryl shikonin （β-HIVS） combined cisplatin on activities of ovarian cancer cell line SKOV3 in vivo and its possible mechanisms. Methods Cells were divided into the blank control group and six β-HIVS groups （2 -30 μmol/L）. Effect of β-HIVS at different concentrations on the activities of ovarian cancer cell line SKOV3 was detected using MTT assay. SKOVs cells were treated with cisplatin （10, 20, and 40 μ mol/L） and β-HIVS （0.25, 1, and 2.5 μmol/L） combined cisplatin. Effect of β-HIVS combined cisplatin on the activities of ovarian cancer cell line SKOV3 was determined by MTT assay. The expression of Bcl-2 and Bax after treated by different concentrations of β-HIVS was detected by Western blot. Results The activities of SKOVs were inhibited by different con- centrations of β-HIVS dose-dependently. The 50% inhibition rate （ IC50） was 7.37 μmol/L. There was sta- tistical difference in IC50 between each concentration β-HIVS group and the blank control group （P 〈0.05）. There was statistical difference in IC50 between the β-HIVS （1 and 2.5 μmol/L） combined cisplatin groups and the cisplatin group （P 〈0.05, P 〈0.01 ）. The synergistic effect on β-HIVS showed dose-dependent manner. Results of Western blot showed β-HIVS at different concentrations （5, 7.5, and 10 μmol/L） could obviously up-regulate the expression level of Bax protein and inhibit the expression level of Bcl-2protein, showing statistical difference when compared with the control group （P 〈0.01 ）. Conclusions β- HIVS could obviously inhibit in vitro growth of SKOVs in a dose-dependent manner. With the range of con- centration, β-HIVS showed synergetic effect with cisplatin. Besides, along with increasing β-HIVS concen- trations, the synergetic effect was more significant. The synergetic effect might accelerate the apoptosis of SKOV3 through up-regulating Bax expression and inhibiting Bcl-2 expression.