百里醌对体内外乳腺癌细胞生长和凋亡的作用  

Anti-proliferation and Apoptosis Induction Effects of Thymoquinone on Human Breast Cancer in vitro and in vivo

作　　者：封伟亮[1] 杨红健[1] 谢尚闹[1] 

机构地区：[1]浙江省肿瘤医院乳腺外科,杭州310022

出　　处：《医学研究杂志》2013年第6期94-97,共4页Journal of Medical Research

基　　金：浙江省卫生厅基金资助项目(2008A122)

摘　　要：目的研究百里醌在体内外对人乳腺癌细胞株MCF-7生长和凋亡的影响,并探讨其作用机制。方法采用四甲基偶氮唑蓝(MTT)法检测细胞增殖;AnnexinⅤ-FITC/PI双染法检测细胞凋亡;Western blot检测survivin以及半胱天冬酶(pro-caspase 3、pro-caspase 8、pro-caspase 9)蛋白的表达,并利用活力检测试剂盒检测caspase 3、caspase 8、caspase 9的活力;建立裸鼠乳腺癌皮下移植瘤模型,原位末端标记(TUNEL)法检测肿瘤组织细胞的凋亡情况;采用免疫组化方法检测肿瘤组织中ki-67、survivin和caspase 3的表达。结果百里醌对体外MCF-7细胞有明显的增殖抑制和凋亡诱导作用;百里醌可下调MCF-7细胞中survivin、pro-caspase 3和pro-caspase 8的表达,但对pro-caspase 9的表达无明显影响;活力测定结果进一步显示,百里醌作用MCF-7细胞后,caspase 3和caspase 8被激活,而对caspase 9无明显影响;在体内试验中,百里醌可抑制裸鼠皮下移植瘤生长,同时诱导体内乳腺癌细胞凋亡,另外百里醌还可分别下调和上调肿瘤组织细胞中survivin和caspase 3的表达。结论百里醌对体内外乳腺癌细胞有凋亡诱导作用,死亡配体途径可能是百里醌诱导乳腺癌细胞凋亡的主要机制之一。Objective To investigate the effect of thymoquinone on the proliferation and apoptosis of breast cancer in vitro and in vivo and explore its mechanism. Methods Human breast carcinoma MCF -7 cell line was cultured in vitro, pretreatment MCF -7 cells with different concentrations of thymoquinone and blank control group, proliferation of cells was detected by methyl thiazolyl tetrazolium （MTT） assay. Cell apoptosis was determined by flow cytometry. Western blotting was used to detect the expression of Survivin, pro - caspase 3 and pro - caspase 8 in breast cancer cells. The activity of caspase 3, caspase g and caspase 9 were determined with caspase activity assay kit. Human breast carcinoma MCF- 7 cells were inoculated subcutaneously into the left armpit of nude mice. The mice（ n = 20） were then randomly divided into two groups： the control group and the thymoquinone group. At the end of the study, tumor tissues were collected for analysis. Cell apoptosis was determined by TUNEL assay. Immunohistochemistry was used to detect the positive expression of ki - 67, Survivin and caspase 3 in the xenograft tumors. Results The proliferation of breast cancer cells was inhibited significantly by thymoquinone in a dose - depentent manner. Apoptosis rate induced by the thymoquinone was markedly highter than that of control. The proteins of Survivin, easpase 3 and caspase 8 were decreased in a dose - dependent manner after treated with thymoquinone, whereas treatment of these cells with thymoquinone had no change in proeaspase - 9 level. The activity of caspase 3 and caspase 8 in MCF - 7 cells were increased with the increasing concentration of thymoquinone, but caspase 9 had no changes. Inhibition of tumor growth was demonstrated after treatment with thymoquinone. The weight of tumors were significantly decreased in comparison with those of control group（P 〈0.05）. Apoptosis occurred more frequently in the thymoquinone group than in the control. Compared with the control, treated with thymoquinone significantly

关 键 词：乳腺癌 百里醌 死亡配体途径 

分 类 号：R979.1[医药卫生—药品]