凝血因子Ⅻ刺激外周血单核细胞对卵巢癌细胞侵袭能力的影响

摘要

目的 探索凝血因子Ⅻ刺激正常人外周血单核细胞(monocytes,MOs)后对卵巢癌细胞侵袭能力的影响.方法 正常女性外周血来源MOs在体外经Ⅻ因子刺激12h后,以卵巢癌患者腹水来源的肿瘤相关巨噬细胞(TAM)为阳性对照,通过印度墨汁吞噬实验检测经Ⅻ因子刺激后,MOs吞噬功能的变化;检测经Ⅻ因子刺激后MOs培养上清对卵巢癌细胞系ES-2,SKOV-3,HO-8910体外侵袭能力的影响;采用AP-1、STAT、Inflammation-1、Oncogene-3家族转录因子试剂盒筛选Ⅻ因子可能通过何种细胞信号转导途径激活MOs.结果 (1)卵巢癌腹水来源的TAM和经Ⅻ因子刺激后MOs吞噬印度墨汁的比例分别为0.66±0.04和0.36±0.03,显著高于外周血来源的MOs(0.27±0.018)(P<0.01,P=0.033).(2)卵巢上皮性癌细胞ES-2在MOs+Ⅻ因子组上清为趋化诱导物时侵袭细胞数目为110.9±5.046,略少于腹水组(157.5±14.86),但显著高于使用普通培养基作为趋化诱导物的阴性对照组(39.14±10.91),在MOs+Ⅻ因子组上清中添加Ⅻ抑制剂西妥昔单抗后,侵袭细胞数量减少为42.43±9.097.结论 卵巢癌腹膜内上调的Ⅻ因子可能通过教育和诱导MOs向具有类似TAM特征的M2型巨噬细胞亚型分化,成为加速卵巢癌细胞在腹膜转移的诱导因素之一.%Objective To explore the impact of monocytes (MOs) stimulated by F Ⅻ on the invasion potential of EOC. Methods Monocytes isolated from female healthy donors were culture and stimulated with Factor Ⅻ for 12 h. Tumor associated macrophage (TAM) were derived from ascites of EOC. India ink engulfing test was used to detect phagocytic capacity of MOs stimulated by F Ⅻ. Invasive potential of EOC, including ES-2, SKOV-3, HO-8910 cells were investigated by Matrigel invasion assay. The signaling pathway involved in MOs activated by F Ⅻ were measured by Transfactor family Kits, including AP-1 ,STAT, Inflammation 1 and Oncogene 3. Results (1) The percentage of engulfing Indian ink were 0.66 s0.04 in TAM and O. 36 s0.03 in monoeytes stimulated by FⅫ, which were siguificanfly higher than that mono-cytes from healthy donors(0.27±0.018) (P < 0.01 and P = 0.033). (2) ES-2 cells were shown more invasive cell number when the supematant from MOs+ Ⅻ was used as chemo-attractant (110.9±5.046), fewer than that of ascites group (157.5±14.86), but significantly more than that of negative control group in which cells were cultured with usualmedium (39.14±10. 91). When adding Centuximab into supernatants, decreased invasive cell numbers (42.43±9.097) were observed. Conclusion FⅫ might educate MOs differentiation toward TAM like cells, thus accelerated me-tastasis and invasion of EOC in the peritoneum.