摘要
目的 探讨白细胞介素6（IL-6）诱导卵巢上皮性癌（卵巢癌）细胞对化疗药物产生耐药的机制及相关信号传导通路.方法 应用外源性重组IL-6短期处理或将正义IL-6基因（ssIL-6）稳定转染至A2780细胞（不表达IL-6但表达其受体、对顺铂和紫杉醇敏感）,同时将反义IL-6基因（asIL-6）稳定转染至SKOV3细胞（高表达IL-6及其受体、对顺铂和紫杉醇耐药）,应用四甲基偶氮唑蓝（MTT）比色法、逆转录PCR（RT-PCR）技术及蛋白印迹法（western blot）观察IL-6是否影响卵巢癌细胞对顺铂和紫杉醇的敏感性,并对可能的信号传导通路进行研究.结果 （1）外源性重组IL-6处理A2780细胞后,对顺铂和紫杉醇的耐药倍数分别为6.25和7.31倍;ssIL-6转染A2780细胞后（内源性过表达IL-6）,IL-6低、中、高表达细胞对顺铂的耐药倍数分别为7.13、7.47和8.34倍,对紫杉醇的耐药倍数分别为7.61、8.27和10.70倍;而asIL-6转染SKOV3细胞后（抑制内源性IL-6表达）,IL-6中、高抑制表达细胞对顺铂的耐药倍数分别为0.15和0.10倍,对紫杉醇的耐药倍数分别为0.10和0.08倍,可恢复SKOV3细胞对顺铂和紫杉醇的敏感性.（2）外源性IL-6作用后,上调了A2780细胞中耐药相关基因MDR1和GST-π以及凋亡抑制基因bcl-2、bcl-xL和XIAP mRNA和蛋白的表达;与此相一致,ssIL-6转染的A2780细胞中MDR1、GST-π及bcl-2、bcl-xL、XIAP mRNA和蛋白的表达水平增加,而asIL-6转染的SKOV3细胞则明显降低.（3）有丝分裂原活化的蛋白激酶-细胞外信号调节激酶（MEK）抑制剂--PD98059和磷脂酰肌醇3激酶（PI3K）抑制剂--wortmannin能分别阻断IL-6诱导的卵巢癌细胞细胞外信号调节激酶（ERK）和蛋白激酶B（Akt）的活化作用,且两者均能阻断IL-6诱导的卵巢癌细胞对顺铂和紫杉醇产生的耐药,细胞生长明显减少.结论 IL-6诱导的卵巢癌细胞化疗耐药很可能与其上调卵巢癌细胞耐药相关基因MDR1、GST-π和凋亡抑制基因bcl-2、bcl-xL、XIAP的表达以及活化MEK/ERK和PI3K/Akt信号传导通路有关.
Objective To study the mechanism of chemotherapy resistance caused by interleukin-6 （IL-6） in ovarian cancer cells and its related signal pathways. Methods Ovarian cancer cell lines A2780（IL-6 receptor positive, while non-IL-6-expressing and cisplatin/paclitaxel-responsive） and SKOV3 cell lines（ overexpressing of IL-6 receptor and IL-6 and cisplatin/paclitaxel-resistant） were suitable models for this study. The effect of exogenous （a short period of treatment with recombination IL-6） and endogenous IL-6（by transfecting with plasmid encoding for sense IL-6 ） in A2780 cells or deleting of endogenous IL-6expression in SKOV3 cells （by transfecting with plasmid encoding for antisense IL-6） on the sensitivity to cisplatin and paclitaxel was investigated. Meanwhile, the mechanism of chemotherapy resistance caused by IL-6 in ovarian cancer cells and its related signal pathways were also analyzed. Results We found that both exogenous and endogenous IL-6 induce cisplatin and paclitaxel resistance in non-IL-6-expressing A2780 cells （the resistance multiple to cisplatin and paclitaxel was： exogenous, 6. 25 and 7.31; endogenous, 7. 13 -8. 34 and 7. 61 - 10. 70）, while deleting of endogenous IL-6 expression in IL-6-overexpressing SKOV3 cells promotes its sensitivity to anticancer drugs （ the resistance multiple to cisplatin and paclitaxel was 0. 15 and 0. 10, 0. 10 and 0. 08）. IL-6 significantly up-regulated the expression levels of mRNA and protein of drug resistance-associated genes, MDR1 and GST-π, and apoptosis-inhibiting genes, bcl-2, bcl-xL and XIAP in a dose-dependent manner in A2780 cells. In accordance with this finding, the mRNA and protein levels of MDR1 and GST-π enhanced in sense IL-6-transfected A2780 cells, and reduced in antisense IL-6-transfected SKOV3 cells compared with the corresponding parental and control vector-transfected cells, which had no difference. It was found that PD98059 [ mitogen-activated protein kinase-extracellular signalregulated kinase （MEK） inhibitor ] and wortmannin [ phosphatidylinositol 3-kinase （PI3K） inhibitor ]significantly antagonized IL-6-induced phosphorylation of extracellular signal-regulated kinase （ ERK ） and protein kinase B （Akt）, respectively, and both of them blocked IL-6-induced cisplatin and paclitaxel resistance and the inhibitory effects of PD98059 and wortmannin were dependent on its concentration.Conclusions These data suggest that IL-6-induced chemoresistance may be associated with increase of both drug resistance-associated genes （ MDR1 and GST-π） and apoptosis-inhibiting genes （ bcl-2, bcl-xL and XIAP）, and activation of MEK/ERK and PL3K/Akt. Therefore, modulation of IL-6 expression or its related signaling pathway may be a promising strategy of treatment for drug-resistant ovarian cancer.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2010年第9期691-698,共8页
Chinese Journal of Obstetrics and Gynecology
基金
基金项目：天津市应用基础及前沿科技研究计$11（08JCYBJC06900）
中国博士后科学基金（20080441340）
武警医学院科学技术研究重点项目（WJZ2007.1）
关键词
卵巢肿瘤
抗药性
肿瘤
白细胞介素6
信号传导
Ovarian neoplasms
Drug resistance, neoplasm
Interleukin-6
Signal transduction
