异甘草素诱导人卵巢癌细胞株SKOV3凋亡及自噬的研究

Pro-apoptosis and pro-autophagy effect of isoliquiritigenin on human ovarian cancer cell lines SKOV3

目的观察异甘草素(ISL)对人卵巢浆液性乳头状腺癌细胞株SKOV3体外增殖的抑制作用,并探讨其可能存在的分子机制。方法四甲基偶氮唑蓝(MTT)比色法观察ISL对SKOV3细胞生长的抑制作用;吖啶橙/溴化乙锭双荧光染色后观察细胞形态变化;流式细胞术AnnexinⅤ-FITC/PI双染检测ISL对细胞凋亡的影响;Western blot检测凋亡相关蛋白Bcl-2、Bax及自噬相关蛋白Beclin1、LC3的表达水平。结果 ISL能够抑制SKOV3细胞的增殖,呈时间、剂量依赖性;荧光染色后可见细胞核变圆,碎裂,染色质浓缩;流式细胞仪检测ISL(5、10、20μg/mL)作用于SKOV3细胞48h后,凋亡率明显高于ISL 0μg/mL组,差异具有统计学意义(P<0.05);Western blot结果显示ISL(5、10、20μg/mL)作用于SKOV3细胞48h后Bcl-2蛋白的表达下调,Bax、Beclin1、LC3蛋白的表达上调,差异有统计学意义(P<0.05)。结论 ISL对人卵巢癌SKOV3细胞具有明显的抑制增殖和诱导凋亡的作用,其抗体外肿瘤机制可能与激活线粒体凋亡通路及自噬通路相关。

Objective To investigate the proliferation‐inhibition activity ,pro‐apoptosis and pro‐autophagy of isoliquiritigenin on ovarian cancer cell lines SKOV3 in vitro ,and explore its mechanism .Methods The cytotoxic effect of Isoliquiritigenin（ISL） on the SKOV3 cells was tested by the means of MTT .Cell morphology was observed with invert microscope after AO/EB staining .The apoptosis of the SKOV3 cells was detected with flow cytometer .The expression levels of Bax ,Bcl‐2 ,Beclin1 and LC3 were detected by Western‐blotting .Results Different concentrations of ISL（5 ,10 ,20 μg/mL） showed the inhibitory effect on SKOV3 cells and presented the characteristic morphological changes of apoptosis under inverted microscope .The FCM analysis showed that the ap‐optosis rate of the SKOV3 cells was increased after treatment of ISL accompanied a concentration increment and a prolongation of the culture time（P〈 0 .05） .Western‐blotting result showed that the expression levels of Bax ,Beclin1 and LC3 were increased in SKOV3 treated with ISL （5 ,10 ,20 μg/mL） ,while the expression level of Bcl‐2 was significantly decreased （P〈0 .05） .Conclusion ISL could inhibit the proliferation of the SKOV3 cells in vitro and induce apoptosis ,and that maybe associate with the mitochon‐drial pathway of apoptosis and autophagy pathway .