CAJ | 학술논문

目的探讨百里醌联合阿霉素对人骨肉瘤细胞生长的影响及其机制.方法应用细胞计数试剂盒(CCK-8)法检测细胞存活率；流式细胞术检测细胞凋亡；Western blot检测bax及B淋巴细胞/白血病-2(bcl-2)的表达；建立裸鼠骨肉瘤皮下移植瘤模型,计算抑瘤率,免疫组织化学法检测裸鼠肿瘤组织中细胞核增殖抗原(Ki-67)、bax及bcl-2的表达.结果阿霉素或百里醌单独及阿霉素联合百里醌作用骨肉瘤细胞后,细胞存活率分别为(78.94±8.75)％、(67.41±5.26)％和(38.81 ±3.58)％,细胞早期凋亡率分别为(8.85±1.25)％、(11.26±2.51)％和(27.15±3.25)％,联合用药组细胞存活率均低于各单药组和对照组(P＜0.05),联合用药组细胞凋亡率均高于其他各组(P＜0.05)；阿霉素联合百里醌作用于骨肉瘤细胞后,骨肉瘤细胞中bax蛋白表达明显增加,丽bcl-2的表达显著减少；体内实验显示百里醌组、阿霉素组和联合用药组抑瘤率分别为37.51％、48.37％和66.85％,联合用药组与各单药组及对照组比较,差异有统计学意义(P＜0.05)；与对照组比较,联合用药组裸鼠肿瘤组织中Ki-67和bel-2的阳性表达明显减弱,而bax的表达明显增强,差异均有统计学意义(P＜0.05).结论百里醌联合阿霉素可明显增强阿霉素对体外及体内骨肉瘤细胞的生长抑制作用,该作用可能通过下调bcl-2及上调bax表达而实现.
목적 탐토백리곤연합아매소대인골육류세포생장적영향급기궤제.방법 응용세포계수시제합(CCK-8)법검측세포존활솔；류식세포술검측세포조망；Western blot검측bax급B림파세포/백혈병-2(bcl-2)적표체；건립라서골육류피하이식류모형,계산억류솔,면역조직화학법검측라서종류조직중세포핵증식항원(Ki-67)、bax급bcl-2적표체.결과 아매소혹백리곤단독급아매소연합백리곤작용골육류세포후,세포존활솔분별위(78.94±8.75)％、(67.41±5.26)％화(38.81 ±3.58)％,세포조기조망솔분별위(8.85±1.25)％、(11.26±2.51)％화(27.15±3.25)％,연합용약조세포존활솔균저우각단약조화대조조(P＜0.05),연합용약조세포조망솔균고우기타각조(P＜0.05)；아매소연합백리곤작용우골육류세포후,골육류세포중bax단백표체명현증가,려bcl-2적표체현저감소；체내실험현시백리곤조、아매소조화연합용약조억류솔분별위37.51％、48.37％화66.85％,연합용약조여각단약조급대조조비교,차이유통계학의의(P＜0.05)；여대조조비교,연합용약조라서종류조직중Ki-67화bel-2적양성표체명현감약,이bax적표체명현증강,차이균유통계학의의(P＜0.05).결론 백리곤연합아매소가명현증강아매소대체외급체내골육류세포적생장억제작용,해작용가능통과하조bcl-2급상조bax표체이실현.
Ohjective To investigate the effect of thymoquinone on the sensitization of osteosarcoma to doxorubicin as well as its mechanisam.Methods After human osteosarcoma SaOS-2 cells were treated with thymoquinone,doxorubicin,and thymoquinone combined with doxorubicin,the cellular proliferation was detected by using cell counting Kit-8 (CCK-8) assay.The flow cytometry (FCM) was used to determine apoptosis of SaOS-2 cells.Western blotting was used to detect the protein expression of B lymphocytes/leukemia-2 (bcl-2) and bax.In vivo antitumor effects of thymoquinone and doxorubicin were assessed using SaOS-2 xenograft in BALB/C nude mice model.Immunohistochemistry was used to detect the positive expression of proliferation cell nuclear antigen (Ki-67),bcl-2 and bax in the xenograft tumors.Results The viability rate of SaOS-2 cells in thymoquinone group,doxorubicin group and thymoinone + doxorubicin group was (78.94 ± 8.75)％,(67.41 ± 5.26)％ and (38.81 ± 3.58)％,respectively.The apoptosis rate in thymoquinone group,doxorubicin group and thymoinone + doxorubicin group was (8.85 ± 1.25) ％,(11.26 ± 2.51) ％ and (7.15 ± 3.25) ％,respectively.The expression of bax and bcl-2 was up-regulated and down-regulated,respectively in SaOS-2 cells after co-treatment of thymoquinone plus doxorubicin.Thymoquinone or doxorubicin alone caused 37.51％ and 48.37％ reduction in tumor weight,respectively,but administration of thymoquinone combined doxorubicin caused 66.85％ reduction in tumor weight.As compared with control group and doxoubicin group,thymoquinone + doxorubicin significantly increased the expression of bax in tumor tissues,but the positive expression of Ki-67 and bcl-2 was decreased.Conclusion Thymoquinone could potentiate the growth inhibition of osteosarcoma induced by doxorubicin both in vitro and in vivo,which may be related to up-regulation of bax and down-regulation of bcl-2.