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目的分析比较4种常见的人上皮性卵巢癌细胞系IL-6、IL-8分泌与其化疗敏感性及部分耐药相关基因和凋亡抑制基因表达的关系,为今后研究IL-6、IL-8诱导卵巢癌细胞对化疗药物产生耐药的机制奠定基础。方法IL-6、IL-8及其受体的表达采用RT-PCR、ELISA及免疫印迹技术进行检测,卵巢癌细胞对顺铂和紫杉醇的敏感性采用MTT试验进行分析,耐药相关基因和凋亡抑制基因的表达则采用RT-PCR进行测定。结果1)4种卵巢癌细胞除A2780细胞外均组成性分泌IL-6和IL-8,二者转录水平与其蛋白水平基本一致。2)4种卵巢癌细胞均表达IL-6受体和IL-8受体。3)4种卵巢癌细胞对顺铂和紫杉醇的敏感性不同,其中A2780细胞最敏感,ES-2细胞次之,CAOV-3和SKOV-3细胞则有不同程度的耐药性。4)部分耐药相关基因和凋亡抑制基因在A2780和ES-2细胞中的表达水平均较低,而在CAOV-3和SKOV-3细胞中的表达水平均较高。结论卵巢癌细胞IL-6、IL-8的自分泌水平(尤其是IL-6的水平)与其对顺铂和紫杉醇的敏感性基本呈负相关趋势,与其部分耐药相关基因和凋亡抑制基因的表达水平相一致。
Abstract：Objective To investigate the relationship of IL-6 and IL-8 secretion in four epithelial ovarian cancer cell lines(A2780,CAOV-3,SKOV-3 and ES-2) with their sensitivity to chemotherapy or the expression of several drug resistance-associated genes and apoptosis-inhibiting genes.In an attempt to provide a basis for further investigating the mechanism of chemotherapy resistance caused by IL-6 and IL-8 in ovarian cancer cells.Methods Reverse transcription polymerase chain reaction(RT-PCR),enzyme linked immunosorbent assay(ELISA) and Western blot analysis were performed to analyze the expression of IL-6,IL-8,and their receptors.MTT assay was carried out to examine the sensitivity to cisplatin and paclitaxel in ovarian cancer cells.The expressions of drug resistance-associated genes and apoptosis-inhibiting genes were detected by RT-PCR.Results Four ovarian cancer cells,except A2780 cells,constitutively expressed IL-6 and IL-8.The mRNA levels of IL-6 and IL-8 resembled their respective protein levels in four ovarian cancer cells.Four cell lines all expressed IL-6R,gp130,IL-8RA,and IL-8RB.The sensitivity to cisplatin and paclitaxel in four ovarian cancer cells was different.A2780 cells were the most sensitive cells and ES-2 cells were less,whereas CAOV-3 and SKOV-3 cells were drug-resistant.The expression levels of drug resistance-associated genes and apoptosis-inhibiting genes were lower in A2780 and ES-2 cells,as compared with CAOV-3 and SKOV-3 cells.Conclusion Autocrine levels of IL-6 and IL-8(especially IL-6) in epithelial ovarian cancer cell lines are inversely associated with their sensitivity to cisplatin or paclitaxel,and are in agreement with the expression levels of several drug resistance-associated genes and apoptosis-inhibiting genes in these cells.
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基本信息:
中图分类号:R737.31
引用信息:
[1]王越,牛秀珑,郭小芹,等.卵巢癌细胞IL-6、IL-8分泌与其化疗敏感性及耐药相关基因和凋亡抑制基因表达关系的初步探讨[J].免疫学杂志,2009,25(04):436-441.
基金信息:
天津市自然科学基金项目(08JCYBJC06900);; 武警医学院科学技术研究重点项目(WJZ2007-1);; 博士科研启动金项目(WBS2007-6)
2009-07-15
2009-07-15