卵巢癌细胞IL-6、IL-8分泌与他莫西芬敏感性及MAPK、Akt活性和ER特异性位点磷酸化水平的关系

Relationship of IL-6 and IL-8 secretion in epithelial ovarian cancer cell lines with their sensitivity to tamoxifen as well as MAPK,Akt and estrogen receptor phosphorylation

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分泌与他莫西芬(TAM)敏感性及MAPK、Akt活性和雌激素受体(ER)特异性位点磷酸化水平的关系,探讨IL-6、IL-8诱导卵巢癌细胞对内分泌治疗产生耐药的机制。方法:RT-PCR及ELISA法检测IL-6、IL-8的表达,MTT法分析卵巢癌细胞对TAM的敏感性,免疫印迹法测定MAPK、Akt活性及ER特异性位点的磷酸化水平。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,其耐药倍数分别为4.99、3.76和2.66;(3)4种卵巢癌细胞的MAPK、Akt活性存在差异,CAOV-3细胞的二者活性最强,SKOV-3细胞的MAPK活性弱于ES-2细胞,而其Akt活性则强于ES-2细胞,A2780细胞未检测到二者有活性;(4)ER的第167位丝氨酸(Ser167)在四种卵巢癌细胞中均存在不同程度的磷酸化,ER的第118位丝氨酸(Ser118)除A2780细胞外亦存在不同程度的磷酸化。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MAPK、Akt活性和ER特异性位点(Ser167、Ser118)的磷酸化水平相一致。

AIM：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 tamoxifen（TAM）as well as MAPK,Akt and estrogen receptor（ER）phosphorylation,and to explore the mechanism of endocrine therapy resistance caused by IL-6 and IL-8 in ovarian cancer cells.METHODS：Reverse transcription polymerase chain reaction（RT-PCR）and enzyme linked immunosorbent assay（ELISA）were performed to analyze the expression of IL-6 and IL-8.MTT assay was carried out to examine the response of ovarian cancer cells to TAM.Western blot was used to detect phosphorylated MAPK,Akt and ER.RESULTS：Except A2780 cells,three other ovarian cancer cells constitutively expressed IL-6 and IL-8.The mRNA levels of IL-6 and IL-8 correlated with their protein levels in four ovarian cancer cells.The four ovarian cancer cells showed different response to TAM.A2780 cells was the most responsive,whereas CAOV-3,SKOV-3 and ES-2 cells were TAM-resistant to a different degree.There was a notable difference in phosphorylated MAPK,Akt and ER（serine 118 and 167）among the four ovarian cancer cells.CONCLUSION：Autocrine production of IL-6 and IL-8 in epithelial ovarian cancer cell lines is inversely associated with cell response to TAM,and positively associated with phosphorylated MAPK,Akt and ER.