卵巢癌细胞IL-6、IL-8分泌与其他莫西芬敏感性及ER亚型及p160共激活因子表达关系的初步探讨

The relationship of IL-6 and IL-8 secretion in epithelial ovarian cancer cell lines with their sensitivity to tamoxifen and the expression of estrogen receptor isoforms and p160 coactivators

目的分析比较4种常见的人上皮性卵巢癌细胞系(A2780、CAOV-3、SKOV-3和ES-2)IL-6、IL-8分泌与其他莫西芬(Tamoxifen,TAM)敏感性及雌激素受体(estrogen receptor,ER)亚型及p160共激活因子表达的关系,为今后研究IL-6、IL-8诱导卵巢癌细胞对内分泌治疗产生耐药的机制奠定基础。方法 IL-6、IL-8的表达采用RT-PCR及ELISA法进行检测,卵巢癌细胞对TAM的敏感性采用MTT试验进行分析,ERα、ERβ及p160共激活因子的表达采用免疫印迹和RT-PCR技术进行检测。结果 4种卵巢癌细胞除A2780细胞外均组成性分泌IL-6和IL-8;4种细胞对TAM的敏感性不同,其中A2780细胞最敏感,CAOV-3、SKOV-3和ES-2细胞则有不同程度的耐药性(耐药倍数分别为4.98、3.75和2.66);这些细胞均不同程度地表达ERα,其中A2780细胞较低,CAOV-3和SKOV-3细胞较高,ES-2细胞居中;ERβ的表达情况则与ERα恰好相反;3种p160共激活因子mRNA的表达水平均以A2780细胞为最低,而ES-2、SKOV-3和CAOV-3细胞则依次升高。结论卵巢癌细胞IL-6、IL-8的自分泌水平与其对TAM的敏感性呈负相关,与其ERβ(而非ERα)及3种p160共激活因子的表达水平相一致。

We investigated 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 tamoxifen （TAM） and the expression of estrogen receptor （ER） isoforms and p160 coactivators,in an attempt to provide a basis for further investigating the mechanism of endocrine therapy resistance caused by IL-6 and IL-8 in ovarian cancer cells.Reverse transcription polymerase chain reaction （RT-PCR） and enzyme linked immunosorbent assay （ELISA） were performed to analyze the expression of IL-6 or IL-8.MTT assay was carried out to examine the sensitivity to TAM in ovarian cancer cells.The expressions of ER isoforms and p160 coactivators were detected by RT-PCR and Western blotting.Four kinds of ovarian cancer cells constitutively expressed IL-6 and IL-8 except A2780 cells.The sensitivity to TAM in four kinds of ovarian cancer cells was different.A2780 cells were the most sensitive,whereas CAOV-3,SKOV-3,and ES-2 cells were TAM-resistant at the different degree.Lower level of ERα was observed in A2780 cells,higher level was found in CAOV-3 and SKOV-3 cells,while moderate level was detected in ES-2 cells.The ERβ expression was opposite to the ERα expression.A gradual up-regulation of three p160 coactivators mRNA was detected in A2780,ES-2,SKOV-3 and CAOV-3 cells.We concluded that autocrine production levels of IL-6 and IL-8 in epithelial ovarian cancer cell lines are inversely associated with their sensitivity to TAM,and are in agreement with the expression of ERα （but not ERβ） and p160 coactivators in these cells.