龙胆苦苷对卵巢癌细胞株SKOV3氧化应激损伤和细胞凋亡的影响及机制研究  

Effect and Mechanism of Gentiopicroside on Oxidative Stress and Apoptosis in Ovarian Cancer Cell Line SKOV3

作　　者：陈杰 佟玲[1] 朱亦峰 CHEN Jie;TONG Ling;ZHU Yifeng(Department of Obstetrics and Gynecology, Huludao Central Hospital, Huluclao, Liaoning, 125000, China)

机构地区：[1]葫芦岛市中心医院妇产科,辽宁省葫芦岛市125000

出　　处：《医学分子生物学杂志》2019年第2期125-130,共6页Journal of Medical Molecular Biology

基　　金：辽宁省自然科学基金(No.2014021104).

摘　　要：目的探讨龙胆苦苷(Gentiopicroside,GP)对人卵巢癌SKOV3细胞氧化应激和细胞凋亡的影响及相关的机制方法细胞计数试剂盒8(cell counting kit8,CCK-8)检测龙胆苦苷对卵巢癌SKOV3细胞生长的作用;流式细胞术检测细胞凋亡情况;蛋白质印迹检测龙胆苦苷对细胞凋亡相关蛋白表达的影响;试剂盒检测超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathion peroxidase,Gpx)的活性和丙二醛(malondialdehyde,MDA)的含量;蛋白质印迹检测Nrf2、H0-1和NQO-1的表达水平;龙胆苦甘添加至sh-Nrf2转染的SKOV3细胞后,检测SOD的含量和细胞凋亡情况匸结果与SKOV3组相比,龙胆苦苷加药组细胞增殖倍数明显降低;发生凋亡的细胞比率明显增加;Bcl-2的表达水平显著降低,Bax的表达水平显著上升;SOD、Gpx的活性显著降低.MDA的含量显著上升;NF-E2相关因子2(NF-E2 related factor2,NrO)、血红素氧合酶(heme oxygenase,HO-1)和NAD(P)H脱氢(醍)酶[NAD(P)H dehydrogenase (quinone1).NQO-1]的表达水平均显著降低;且随着龙胆苦苷浓度的增加,效果越明显shKNA-Nrf2转架SKOV3细胞后,细胞中Nrf2的表达水平显著降低.龙胆苦甘添加到shRNA转染的SKOV3细胞后,细胞中SOD的活性没有明显变化,凋讥细胞比率没有明显变化结论龙胆苦苷促进卵巢癌SKOV3细胞氧化应激和细胞凋亡,其作用机制的Nrf2通路的抑制相关。Objective To investigate the effects and mechanism of gentiopicroside on oxidative stress and apoptosis of hunian ovarian cancer SKOV3 cells. Methods CCK-8 was used to detect the effect of gentiopicroside on the growth of ovarian cancer SKOV3 cells. Flow cytometry was used to detect apoptosis;Western blotting was used to detect the effect of gentiopicroside on the expression of apoptosis-relatrd proteins and the expression of Nrf2 , HO-1 and NQO-1. Related kits was used to detect the activity of SOD, Gpx and the content of MDA. Gentiopicroside was added to sh-Nrf2 transfected SKOV3 cells to detect SOD content and apoptosis. Results Compared with the SKOV3 group, the gentiopicroside plus group cell proliferation was significantly reduced;the proportion of apoptotic cells increased significantly. The expression level of Bcl-2 decreased significantly and the expression level of Bax increased significantly. The activity of SOD and Gpx decreased significantly,the content of MDA increased significantly. The expression levels of Nrf2 , HO-1 and NQO-1 were significantly reduced. And with the concentration of gentiopicrin increased , the effect was more obvious. After transfection of SKOV3 cells with shRNA-Nrf2, the expression level of NrF2 decreased significantly. After gentiopicroside was added to shRNA-transfected SKOV3 cells, SOD activity and apoptotic cell ratio did not change signific antly. Conclusion Gentiopicroside promotes oxidative stress and cell apoptosis in ovarian cancer SK0V3 cells, which is related to the inhibition of Nrf2 pathway.

关 键 词：卵巢癌 龙胆苦苷 氧化应激 细胞凋亡 Nrf2信号通路 

分 类 号：R737.31[医药卫生—肿瘤]