百里醌联合阿霉素对体内外骨肉瘤生长的影响  

Thymoquinone potentiates antitumor activity of doxorubicin in osteosarcoma in vitro and in vivo

作　　者：沈为栋[1] 陈飞雨[2] 刘岸[1] 尚雷[2] 刘昌戎[1] 彭伟[1] 

机构地区：[1]中南大学湘雅医学院岳阳临床学院,湖南岳阳414000 [2]中南大学湘雅附二医院骨科,湖南岳阳414000

出　　处：《中华实验外科杂志》2013年第2期363-366,共4页Chinese Journal of Experimental Surgery

基　　金：湖南省科技厅资助项目（2011SK3039）

摘　　要：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；Westernblot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，而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bcl-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。Objective To investigate the effect of thymoquinone on the sensitization of osteosarco- ma to doxorubicin as well as its mechanisam. Methods After human osteosarcoma SaOS-2 cells were trea- ted with thymoquinone, doxorubicin, and thymoquinone combined with doxorubicin, the cellular prolifera- tion was detected by using cell counting Kit-8 （ CCK-8 ） assay. The flow cytometry （FCM） was used to de- termine apoptosis of SaOS-2 cells. Western blotting was used to detect the protein expression of B lympho- cytes/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 doxorubicin group, thymoquinone ± doxorubicin significantly increased the expression of bax in tumor tissues, but the positive expression of Ki-67 and bcl-2 was decreased. Conclu- sion 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.

关 键 词：骨肉瘤 阿霉素 百里醌 脱噬作用 BAX 

分 类 号：R73[医药卫生—肿瘤]