CAJ | 학술논문

目的：探讨Bcl-2、Bax在氧化苦参碱联合顺铂抑制卵巢癌生长中的作用机制。方法50只雌性Fish-er344大鼠，腋部皮下接种NuTu-19细胞，建立卵巢癌实体瘤模型，接种成功后，将荷瘤大鼠随机分为5组：模型组、顺铂组（3 mg／kg）、顺铂＋氧化苦参碱（40 mg／kg、80 mg／kg、160 mg／kg）组，每组10只。治疗12 d后，称量肿瘤重量并计算抑制率，流式细胞仪测定肿瘤组织细胞凋亡率。将卵巢癌细胞株SKOV3分别经空白对照组、顺铂组、顺铂＋氧化苦参碱（2┅．5 mg／ml、5．0 mg／ml、10．0 mg／ml ）组作用48 h后，MTT法检测细胞增殖，流式细胞术检测细胞周期及凋亡率， Western blot检测Bcl-2、Bax 蛋白表达。结果氧化苦参碱160 mg／kg、80 mg／kg组能够明显降低肿瘤重量（ P ＜0．05），提高肿瘤抑制率。与顺铂组相比，氧化苦参碱明显提高肿瘤细胞凋亡率，其中氧化苦参碱160 mg／kg组作用最为明显（ P ＜0．05）；与顺铂组相比，氧化苦参碱明显抑制SKOV3细胞增殖，50．mg／ml、10．0 mg／ml组与顺铂组比较，差异有统计学意义（ P ＜0．05），呈浓度依赖性。氧化苦参碱能增强顺铂对细胞周期的作用，与顺铂组相比G0／G1期细胞显著增多（ P ＜0．05），同时顺铂＋氧化苦参碱组较顺铂组细胞凋亡率有所升高，10．0 mg／ml剂量组与顺铂组比较，差异有统计学意义（ P ＜0．05）。氧化苦参碱明显下调Bcl-2蛋白表达，5．0 mg／ml、10．0 mg／ml组与顺铂组比较，差异有统计学意义（ P ＜0．05）；氧化苦参碱明显上调Bax蛋白表达，10．0 mg／ml组与顺铂组比较，差异有统计学意义（ P ＜0．05）。结论氧化苦参碱具有增强顺铂对卵巢癌生长的抑制作用，该作用可能部分通过下调Bcl-2、上调Bax表达和诱导卵巢癌细胞凋亡而实现。
목적：탐토Bcl-2、Bax재양화고삼감연합순박억제란소암생장중적작용궤제。방법50지자성Fish-er344대서，액부피하접충NuTu-19세포，건립란소암실체류모형，접충성공후，장하류대서수궤분위5조：모형조、순박조（3 mg／kg）、순박＋양화고삼감（40 mg／kg、80 mg／kg、160 mg／kg）조，매조10지。치료12 d후，칭량종류중량병계산억제솔，류식세포의측정종류조직세포조망솔。장란소암세포주SKOV3분별경공백대조조、순박조、순박＋양화고삼감（2┅．5 mg／ml、5．0 mg／ml、10．0 mg／ml ）조작용48 h후，MTT법검측세포증식，류식세포술검측세포주기급조망솔， Western blot검측Bcl-2、Bax 단백표체。결과양화고삼감160 mg／kg、80 mg／kg조능구명현강저종류중량（ P ＜0．05），제고종류억제솔。여순박조상비，양화고삼감명현제고종류세포조망솔，기중양화고삼감160 mg／kg조작용최위명현（ P ＜0．05）；여순박조상비，양화고삼감명현억제SKOV3세포증식，50．mg／ml、10．0 mg／ml조여순박조비교，차이유통계학의의（ P ＜0．05），정농도의뢰성。양화고삼감능증강순박대세포주기적작용，여순박조상비G0／G1기세포현저증다（ P ＜0．05），동시순박＋양화고삼감조교순박조세포조망솔유소승고，10．0 mg／ml제량조여순박조비교，차이유통계학의의（ P ＜0．05）。양화고삼감명현하조Bcl-2단백표체，5．0 mg／ml、10．0 mg／ml조여순박조비교，차이유통계학의의（ P ＜0．05）；양화고삼감명현상조Bax단백표체，10．0 mg／ml조여순박조비교，차이유통계학의의（ P ＜0．05）。결론양화고삼감구유증강순박대란소암생장적억제작용，해작용가능부분통과하조Bcl-2、상조Bax표체화유도란소암세포조망이실현。
Objective To investigate the antitumor effects of oxymatrine combined with cisplatin by down-regulating Bcl-2 expression and up-regulating Bax expression in ovarian cancer, and to explore the action effects of Bcl-2 and Bax on ovarian cancer in vitro and in vivo.Methods Fifty female Fisher344 rats were injected with NuTu 19 cells hypodermically to establish solid tumor models with ovarian cancer, then the tumor-bearing rats were randomly divided into 5 groups: model group, cisplatin group (3mg/kg), cisplatin+oxymatrine tumor weights (40mg/kg,80mg/kg,160mg/kg) groups,with 10 rats in each group.After 12-day treatment, the tumor weight was measured and the inhibition rate on tumor was calculated. Apoptosis rate of tumor cells was detected by flow cytometry.The ovarian cancer cell strain-SKOV3 O was divided into three groups:blank control group, cisplatin group, cisplatin +oxymatrine ( 2.5mg/ml, 5.0mg/ml, 10.0mg/ml ) groups, after 48-hour treatment, the cell proliferation was detected by MTT, cell cycle and cell apoptosis rate were measured by flow cytometry (FCM) and the expresson levels of Bcl-2,Bax protein were detected by Western Blot.Results Moreover,inhibition tumor rate was significantly increased ( P <0.05) .As compared with that in cisplatin group, the apoptosis rate of tumor cells was significantly increased in oxymatrine groups,especially in 160mg/kg group ( P <0.05).As compared with that in cisplatin group, the proliferation of SKOV3 cells was significantly inhibited by oxymatrine, especially in 5.0mg/ml group and 10.0mg/ml group ( P <0.05),with a dose-dependent way.As compared with that in cisplatin group, the percentage of cells in G0/G1 phase was significantly increased in oxymatrine groups ( P <0.05),moreover,cell apoptosis rate in cisplatin+oxymatrine groups was increased at some extent,as compared with that in cisplatin group, there was a significant difference between oxymatrine 10.0mg/ml group and cisplatin group ( P <0.05).The expression levels of Bcl-2 protein were decreased by oxymatrine,there were significant differences between oxymatrine 5.0mg/ml group, 10.0mg/ml group and cisplatin group (P<0.05).However the expression levels of Bax protein were increased by oxymatrine,there were significant differences between oxymatrine 10.0mg/ml group and cisplatin group ( P <0.05).Conclusion Oxymatrine can enhance the inhibitory effect of cisplatin on growth of ovarian cancer, which may be completed partly by down-regulating expression of Bcl-2 and up-regulating expression of Bax,moreover, by inducing cell apoptosis of ovarian cancer.