雄激素调节卵巢癌细胞IL-6基因启动子活性的研究  

Effect of androgen on the IL-6 promoter activity in ovarian cardnoma cells

作　　者：王越[1] 杨洁[2] 高燕[3] 李宏钊[2] 笪宇荣[2] 姚智[2] 

机构地区：[1]武警医学院免疫学教研室,天津300070300162 [2]天津医科大学免疫学教研室,300070 [3]天津市中心妇产科医院肿瘤科

出　　处：《中华微生物学和免疫学杂志》2007年第10期883-887,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家973重大基础研究前期研究专项项目(2003CCA04300);国家自然科学基金项目(30471962)

摘　　要：目的探讨雄激素诱导卵巢癌细胞IL-6基因启动子活化的分子机制。方法选择SKOV-3细胞作为研究模型,应用ELISA、RT-PCR及荧光素酶检测技术观察5α-二氢睾酮(5α-dihydrotest- osterone,DHT)对IL-6表达水平及IL-6基因启动子活性的影响,并对DHT诱导IL-6基因启动子活化的机制进行研究。结果(1)DHT可促进SKOV-3细胞IL-6分泌及相应mRNA的表达,该作用可被雄激素受体(AR)阻断剂氟他胺(nutamide,Flu)完全阻断,提示DHT诱导上述作用是AR途径介导的。(2)IL-6本身能以剂量依赖性的方式增强SKOV-3细胞IL-6基因启动子的转录活性,DHT也具有与IL- 6相似的作用。DHT的这种作用可被Flu完全阻断,也可被抗IL-6中和抗体部分阻断。(3)转录因子NF-IL-6、NF-κB及其亚单位p50、p65均可诱导SKOV-3细胞IL-6基因启动子的活性。DHT不改变NF- IL-6介导的IL-6基因启动子的转录活化,而增强NF-κB(p50+p65)及其亚单位p50、p65介导的IL-6基因启动子的转录活化。结论雄激素增强的IL-6基因启动子转录活性是AR途径介导的,同时有部分是通过IL-6的作用而实现。雄激素很可能通过AR与转录因子NF-κB或其亚单位p50和p65之间蛋白-蛋白的相互作用反式激活IL-6基因启动子的活性。Objective To investigate the regulation of androgen in the IL-6 promoter activity in ovarian carcinoma cells. Methods SKOV-3 cells were used as suitable model for this study. ELISA, RT-PCR, and lu- ciferase reporter assay were performed to investigate the effect of 5a-dihydrotestostemne （DHT） on IL-6 expression and IL-6 promoter activity. Furthermore, the underline molecular mechanism of DHT-induced IL-6 promoter activation was investigated. Results DHT （0.1-1000 nmol/L） significantly stimulated IL-6 secretion in SKOV-3 cells （ P 〈 0.001）. The mRNA level of IL-6 was down-regulated at early stage of DHT treatment （24 h）, and then up- regulated in a time-dependent manner. Flutamide （Flu）, an androgen receptor（AR） antagonist, completely inhibited IL-6 expression induced by DHT, which suggested that AR pathway played an important role in the regulation of IL-6 secretion. IL-6 itself enhanced IL-6 promoter activation in SKOV-3 cells in a dose-dependent manner. DHT also had this effect, which was completely blocked by Flu （P 〈 0.01 ）, but partially blocked by IL-6-neutralizing antibody （ P 〈 0.05）. Moreover, we found that NF-IL-6, NF-kB p50, p65, and NF-kB （p50 ＋ 1365 combination） were able to induce IL-6 promoter activity. Yet, DHT did not enhance NF-lL-6-mediated activation of the IL-6 promoter. However, DHT did enhance the induction mediated by NF-kB p50, p65, and NF-kB （p50＋ p65 com- bination）. Conclusion Androgen-induced IL-6 promoter activation is mediated by the AR pathway, and in part through the effect of IL-6. Androgen may mediate trans-activation of IL-6 promoter by the protein-protein interaction between AR and NF-kB or pS0 or p65 subunit of NF-kB.

关 键 词：5α-二氢睾酮 IL-6 IL-6基因启动子 细胞转染 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]