Interleukin-8 is a key mediator of FKBP51-induced melanoma growth, angiogenesis and metastasis.
Interleukin-8 is a key mediator of FKBP51-induced melanoma growth, angiogenesis and metastasis.
复制标题
DOI：
10.1038/bjc.2015.154
复制
发表时间：
2015-05-26
影响因子：
8.8
通讯作者：
Singh S
中科院分区：
文献类型：
--
作者：
Srivastava SK;Bhardwaj A;Arora S;Tyagi N;Singh AP;Carter JE;Scammell JG;Fodstad Ø;Singh S
FKBP51在黑色素瘤中过表达，并影响肿瘤细胞的特性。然而，它在黑色素瘤发病机制和潜在机制中的全面作用(S)仍然不清楚。FKBP51在侵袭性黑色素瘤细胞系中稳定沉默，并在体外和小鼠模型中检测其作用。组织学/免疫组织化学分析证实肿瘤转移、血管生成和中性粒细胞浸润。采用定量逆转录聚合酶链式反应、免疫印迹和/或酶联免疫吸附试验检测基因表达。以荧光素酶为基础的启动子报告基因转录活性和启动子结合活性分别用荧光素酶启动子-报告基因和芯片检测。通过基因沉默或中和抗体治疗实现对IL-8的抑制。沉默FKBP51可减少黑色素瘤的生长、转移、血管生成和中性粒细胞的浸润，并通过抑制细胞株和肿瘤移植瘤中的NF-κB下调IL-8的表达。IL-8抑制显著降低FKPB51过表达细胞的生长、迁移和侵袭力；而IL-8处理部分恢复了FKBP51沉默的黑色素瘤细胞受抑制的表型。FKBP51高表达黑色素瘤细胞条件培养液中白介素8的缺失抑制了内皮细胞的增殖和毛细血管样结构的形成，而白介素8处理则促进了FKBP51沉默黑色素瘤细胞条件培养液中培养的内皮细胞的上述作用。FKBP51促进黑色素瘤的生长、转移和血管生成，而IL-8在这些过程中起关键作用。因此，靶向FKBP51或其上游或下游调控通路可能会导致有效的黑色素瘤治疗策略。
FKBP51 is overexpressed in melanoma and impacts tumour cell properties. However, its comprehensive role in melanoma pathogenesis and underlying mechanism(s) remain elusive. FKBP51 was stably silenced in aggressive melanoma cell lines and its effect examined in vitro and in mouse model. Histological/immunohistochemical analyses were performed to confirm metastasis, angiogenesis and neutrophil infiltration. Gene expression was analyzed by qRT–PCR, immunoblot and/or ELISA. NF-κB transcriptional activity and promoter binding were monitored by luciferase-based promoter-reporter and ChIP assays, respectively. Interleukin (IL)-8 inhibition was achieved by gene silencing or neutralising-antibody treatment. FKBP51 silencing reduced melanoma growth, metastasis, angiogenesis and neutrophil infiltration and led to IL-8 downregulation through NF-κB suppression in cell lines and tumour xenografts. IL-8 inhibition drastically decreased growth, migration and invasiveness of FKPB51-overexpressing cells; whereas its treatment partially restored the suppressed phenotypes of FKBP51-silenced melanoma cells. Interleukin-8 depletion in conditioned medium (CM) of FKBP51-overexpressing melanoma cells inhibited endothelial cell proliferation and capillary-like structure formation, whereas its treatment promoted these effects in endothelial cells cultured in CM of FKBP51-silenced melanoma cells. FKBP51 promotes melanoma growth, metastasis and angiogenesis, and IL-8 plays a key role in these processes. Thus, targeting of FKBP51 or its upstream or downstream regulatory pathways could lead to effective therapeutic strategies against melanoma.