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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(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。
OBJECTIVE: To compare the sensitivity of TAM to IL-6 and IL-8 secretion in four common human epithelial ovarian cancer cell lines (A2780, CAOV-3, SKOV-3 and ES- , Akt activity and estrogen receptor (ER) specific site phosphorylation levels of IL-6, IL-8-induced ovarian cancer cells to endocrine therapy resistance mechanism. Methods: The expressions of IL-6 and IL-8 were detected by RT-PCR and ELISA. The sensitivity of ovarian cancer cells to TAM was analyzed by MTT assay. The phosphorylation of MAPK, Akt and ER-specific sites were determined by Western blotting. RESULTS: (1) IL-6 and IL-8 were constitutively secreted by the four kinds of ovarian cancer cells except A2780 cells. The transcriptional level of the two kinds of ovarian cancer cells was basically the same as that of A2780 cells. (2) The sensitivity of four ovarian cancer cells to TAM was different , Of which A2780 cells were the most sensitive, while CAOV-3, SKOV-3 and ES-2 cells had different degrees of drug resistance with resistance rates of 4.99, 3.76 and 2.66 respectively; (3) MAPK of 4 ovarian cancer cells , Akt activity, the activity of CAOV-3 cells was the strongest, the activity of MAPK of SKOV-3 cells was weaker than that of ES-2 cells, while the activity of Akt was stronger than that of ES-2 cells and not detected by A2780 cells (4) ER serine Ser 167 (Ser167) in all four ovarian cancer cells have varying degrees of phosphorylation, ER serine 118 (Ser 118) in addition to A2780 cells also exist different degrees of phosphorylation . CONCLUSION: The autocrine levels of IL-6 and IL-8 in ovarian cancer cells are negatively correlated with their sensitivity to TAM, which are consistent with their MAPK and Akt activities and phosphorylation levels of ER-specific sites (Ser167 and Ser118).