The matrix metalloproteinase-9 regulates the insulin-like growth factor-triggered autocrine response in DU-145 carcinoma cells
The matrix metalloproteinase-9 regulates the insulin-like growth factor-triggered autocrine response in DU-145 carcinoma cells
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DOI：
10.1074/jbc.274.11.6935
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发表时间：
1999-03-12
影响因子：
4.8
通讯作者：
Martínez-A, C
中科院分区：
文献类型：
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作者：
Mañes, S;Llorente, M;Martínez-A, C
不依赖于雄性激素的人前列腺癌细胞系DU-145在无血清培养基中增殖，产生胰岛素样生长因子(IGF)-I、IGF- iz和IGF-1型受体（IGF- 1r），它们还分泌三种IGF结合蛋白（IGFBP）， IGFBP-2、-3和-4，其中，免疫印迹分析显示IGFBP-3选择性蛋白水解，产生31和19 kDa片段。我们在血清饥饿的DU-145细胞上检测到表面受体结合的IGF-I，它激活IGF-IR并触发有丝分裂信号。然而，用抗igf - i、抗igf - ii或抗igf - i加抗igf - ii单抗孵育DU-145细胞不会抑制DU-145的无血清生长，相反，抗igf - 1r单抗和IGFBP-3单抗抑制DNA合成。IGFBP-3还能改变DU-145细胞周期，降低p34（cdc2）水平和IGF-IR自磷酸化。IGFBP-3的抗增殖活性不是独立于igf的，因为des-(1-3)IGF-I不与IGFBP-3结合，从而逆转了其抑制作用。DU-145还分泌基质金属蛋白酶(MMP)-9，可以以可溶性和膜结合形式检测到。基质金属蛋白酶抑制剂，而不是蛇形蛋白酶抑制剂，可以阻断与IGFBP-3蛋白水解相关的DU-145的DNA合成。过表达MMP-9的反义cDNA可抑制80%的DU-145细胞增殖，而IGF-I可以剂量依赖的方式逆转这种增殖。MMP-9表达的抑制也与IGFBP-3蛋白水解的减少和通过IGF- 1r的信号传导减少有关。我们的数据表明，IGF自分泌环在DU-145细胞中运行，由IGFBP-3特异性调节，其活性可能反过来由IGFBP-3蛋白酶如MMP-9调节。
The androgen-independent human prostate adenocarcinoma cell line DU-145 proliferates in serum-free medium and produces insulin-like growth factors (IGF)-I, IGF-IZ, and the IGF type-1 receptor (IGF-1R), They also secrete three IGF-binding proteins (IGFBP), IGFBP-2, -3, and -4, Of these, immunoblot analysis revealed selective proteolysis of IGFBP-3, yielding fragments of 31 and 19 kDa, By using an anti-IGF-I-specific monoclonal antibody (mAb), we detect surface receptor-bound IGF-I on serum-starved DU-145 cells, which activates IGF-IR and triggers a mitogenic signal. Incubation of DU-145 cells with blocking anti-IGF-I, anti-IGF-II, or anti-IGF-I plus anti-IGF-II mAb does not, however, inhibit serum-free growth of DU-145, Conversely, anti-IGF-1R mAb and IGFBP-3 inhibit DNA synthesis. IGFBP-3 also modifies the DU-145 cell cycle, decreases p34(cdc2) levels, and IGF-IR autophosphorylation. The antiproliferative IGFBP-3 activity is not IGF-independent, since des-(1-3)IGF-I, which does not bind to IGFBP-3, reverses its inhibitory effect. DU-145 also secretes the matrix metalloproteinase (MMP)-9, which can be detected in both a soluble and a membrane-bound form. Matrix metalloproteinase inhibitors, but not serpins, abrogate DNA synthesis in DU-145 associated with the blocking of IGFBP-3 proteolysis. Overexpression of an antisense cDNA for MMP-9 inhibits 80% of DU-145 cell proliferation that can be reversed by IGF-I in a dose-dependent manner. Inhibition of MMP-9 expression is also associated with a decrease in IGFBP-3 proteolysis and with reduced signaling through the IGF-1R, Our data indicate an IGF autocrine loop operating in DU-145 cells, specifically modulated by IGFBP-3, whose activity may in turn be regulated by IGFBP-3 proteases such as MMP-9.