Mu Hai-qi, Yang Sen, Wang Yi-jun, Chen Ying-he
Department of Urology, Wenzhou Medical College, Wenzhou, China.
Zhonghua Yi Xue Za Zhi. 2012 Feb 14;92(6):392-6.
To explore the effects and mechanism of thymoquinone in the growth inhibition of bladder cancer both in vitro and in vivo.
After the treatment of thymoquinone, the cellular proliferation of human bladder cancer cell line BIU-87 was detected by the method of methyl thiazolyl tetrazolium (MTT). Flow cytometry (FCM) was used to determine the cellular apoptosis. And the location of nuclear factor (NF)-κB was identified by the method of immunofluorescent histochemistry. Western blotting was employed to detect the cellular expressions of NF-κB, survivin and XIAP. BIU-87 cells were injected subcutaneously into nude mice to establish a xenograft model. After 2 weeks of implantation, the mice were randomized into 2 groups (n = 8): (a) vehicle alone (control), (b) thymoquinone (5 mg/kg daily by intragastric intubation). All treatments lasted for 2 weeks. At Week 7 post-implantation, the mice were sacrificed and their tumor weights and inhibition rates evaluated. Immunohistochemistry was used to detect the positive expressions of Ki-67, NF-κB and XIAP in tumors.
The proliferation of bladder cancer cells was inhibited significantly by thymoquinone at 20, 40, 80 µmol/L (81.2% ± 4.6%, 72.5% ± 6.5%, 58.4% ± 8.9% vs 100%, all P < 0.05). Apoptosis rate induced by thymoquinone was more significant than that of the control (7.6% ± 1.6%, 11.2% ± 2.1%, 14.3% ± 2.8%vs 1.6% ± 0.5%, all P < 0.05). Immunofluorescent histochemistry showed that thymoquinone could significantly lower the nuclear expression of NF-κB. The expressions of NF-κB and XIAP were down-regulated in BIU-87 cells after the treatment of thymoquinone. But thymoquinone had no effect on the expression of survivin. The final tumor weight showed significant decrease in the test group versus the control group ((0.41 ± 0.12) vs (0.89 ± 0.23) g, P < 0.05). Furthermore, the positive expressions of Ki-67, NF-κB and XIAP decreased in tumors after the administration of thymoquinone.
Thymoquinone exerts anti-tumor effects on bladder cancer both in vitro and in vivo through the down-regulations of NF-κB and its regulated molecules such as XIAP.
探讨百里醌在体外和体内对膀胱癌生长抑制的作用及其机制。
用百里醌处理后，采用甲基噻唑基四氮唑（MTT）法检测人膀胱癌细胞系BIU-87的细胞增殖情况。采用流式细胞术（FCM）测定细胞凋亡情况。通过免疫荧光组织化学方法鉴定核因子（NF）-κB的定位。采用蛋白质印迹法检测NF-κB、生存素和X连锁凋亡抑制蛋白（XIAP）的细胞表达。将BIU-87细胞皮下注射到裸鼠体内建立异种移植模型。植入2周后，将小鼠随机分为2组（n = 8）：（a）单独给予溶剂（对照组），（b）百里醌（每天5 mg/kg，经胃内插管给药）。所有处理持续2周。植入后第7周，处死小鼠并评估其肿瘤重量和抑制率。采用免疫组织化学法检测肿瘤中Ki-67、NF-κB和XIAP的阳性表达。
百里醌在20、40、80 μmol/L时可显著抑制膀胱癌细胞的增殖（分别为81.2%±4.6%、72.5%±6.5%、58.4%±8.9%，而对照组为100%，所有P < 0.05）。百里醌诱导的凋亡率比对照组更显著（分别为7.6%±1.6%、11.2%±2.1%、14.3%±2.8%，而对照组为1.6%±0.5%，所有P < 0.05）。免疫荧光组织化学显示，百里醌可显著降低NF-κB的核表达。百里醌处理后，BIU-87细胞中NF-κB和XIAP的表达下调。但百里醌对生存素的表达无影响。试验组的最终肿瘤重量与对照组相比显著降低（（0.41±0.12） vs （0.89±0.23）g，P < 0.05）。此外，给予百里醌后，肿瘤中Ki-67、NF-κB和XIAP的阳性表达降低。
百里醌通过下调NF-κB及其调控分子如XIAP，在体外和体内对膀胱癌发挥抗肿瘤作用。
