Exogenous SPARC Suppresses Proliferation and Migration of Prostate Cancer by Interacting With Integrin β1
Exogenous SPARC Suppresses Proliferation and Migration of Prostate Cancer by Interacting With Integrin β1
复制标题
DOI：
10.1002/pros.22664
复制
发表时间：
2013-08-01
期刊：
影响因子：
2.8
通讯作者：
Namiki, Mikio
中科院分区：
文献类型：
--
作者：
Shin, Minkyoung;Mizokami, Atsushi;Namiki, Mikio
背景富含半胱氨酸的酸性基质细胞分泌蛋白（matricellular protein secreted protein，MMP）在多种肿瘤的转移和进展中起重要作用。然而，前列腺癌（PCa）中的作用仍然不清楚。为了鉴定前列腺组织中的P450蛋白，使用人前列腺组织芯片进行P450的免疫组织化学分析。为了检测前列腺癌（LNCaP、DU 145和PC-3）和基质细胞中的rsB表达，进行了RT-PCR、蛋白质印迹分析和ELISA。为了揭示外源性AKT在PCa细胞中的功能，在与基质细胞共培养后，通过蛋白质印迹分析证实AKT磷酸化。PCa细胞的增殖和迁移通过加入Na 2SO 4来检测。在免疫沉淀后，通过蛋白质印迹分析证实了β 1和整合素β 1之间的相互作用。与PCa组织相比，正常组织中有较好的表达。ELISA检测结果显示，正常前列腺基质细胞（PrSC）分泌的α-SMA蛋白高于前列腺癌基质细胞（PCaSC）和前列腺癌（PCa）。与PrSC共培养的PCa细胞显示AKT磷酸化比PCaSC更低。PCa细胞与PrSC共培养，PrSC敲除AKT，恢复AKT磷酸化。此外，PCa细胞用顺铂处理导致AKT磷酸化减少。免疫沉淀法显示前列腺癌细胞中整合素β 1与β 1的相互作用。整合素β 1中和抗体可恢复PCa细胞增殖和迁移的抑制作用。间质细胞分泌的减少可能通过限制与整合素β 1相互作用后的AKT磷酸化来影响PCa的进展。(C)2013 Wiley Periodicals，Inc.
BACKGROUND. The matricellular protein secreted protein acidic and rich in cysteine (SPARC) plays an important role on tumor metastasis and progression in several cancers. However, the roles of SPARC in prostate cancer (PCa) remain unclear.METHODS. To identify SPARC protein in prostate tissue, immunohistochemical analysis of SPARC was conducted using human prostate tissue microarray. To detect SPARC expression in prostate cancer (LNCaP, DU145, and PC-3) and stromal cells, RT-PCR, western blot analysis, and ELISA was conducted. To reveal the function of exogenous SPARC in PCa cells, AKT phosphorylation was confirmed by western blot analysis after coculture with stromal cells. Proliferation and migration of PCa cells were examined by addition of SPARC. The interaction between SPARC and integrin beta 1 was confirmed by western blot analysis after immunoprecipitation.RESULTS. SPARC protein was expressed well in normal tissue compared with PCa tissue. ELISA showed high secreted SPARC protein in normal prostate-derived stromal cell (PrSC) compared with PCa-derived stromal cell (PCaSC) and PCa. PCa cells cocultured with PrSC showed reduced AKT phosphorylation more than with PCaSC. PCa cells cocultured with PrSC whose SPARC was knocked-down restored AKT phosphorylation. Moreover, PCa cells treated with SPARC led to reduced AKT phosphorylation. Immunoprecipitation with SPARC revealed interaction of SPARC and integrin beta 1 in PCa cells. Inhibited proliferation and migration of PCa cells by SPARC was restored by integrin beta 1 neutralizing antibody.CONCLUSIONS. Reduced SPARC secretion from stromal cells might affect PCa progression mediating through limiting AKT phosphorylation after interaction with integrin beta 1. (C) 2013 Wiley Periodicals, Inc.