Institut de Pharmacologie de Sherbrooke, Department of Surgery/Urology, Université de Sherbrooke, Sherbrooke, Québec, Canada.
Neoplasia. 2012 Nov;14(11):1032-42. doi: 10.1593/neo.121368.
Better understanding of the distinct and redundant functions of the proprotein convertase (PC) enzyme family within pathophysiological states has a great importance for potential therapeutic strategies. In this study, we investigated the functional redundancy of PCs in prostate cancer in the commonly used androgen-sensitive LNCaP and the androgen-independent DU145 human cell lines. Using a lentiviral-based shRNA delivery system, we examined in vitro and in vivo cell proliferation characteristics of knockdown cell lines for the endogenous PCs furin, PACE4, and PC7 in both cell lines. Of the three PCs, only PACE4 was essential to maintain a high-proliferative status, as determined in vitro using XTT proliferation assays and in vivo using tumor xenografts in nude mice. Furin knockdowns in both cell lines had no effects on cell proliferation or tumor xenograft growth. Paradoxically, PC7 knockdowns reduced in vitro cellular proliferation but had no effect in vivo. Because PCs act within secretion pathways, we showed that conditioned media derived from PACE4 knockdown cells had very poor cell growth-stimulating effects in vitro. Immunohistochemistry of PACE4 knockdown tumors revealed reduced Ki67 and higher p27(KIP) levels (proliferation and cell cycle arrest markers, respectively). Interestingly, we determined that the epidermal growth factor receptor signaling pathway was activated in PC7 knockdown tumors only, providing some explanations of the paradoxical effects of PC7 silencing in prostate cancer cell lines. We conclude that PACE4 has a distinct role in maintaining proliferation and tumor progression in prostate cancer and this positions PACE4 as a relevant therapeutic target for this disease.
更好地理解蛋白水解酶(PC)酶家族在生理病理状态下的独特和冗余功能对于潜在的治疗策略具有重要意义。在这项研究中,我们研究了常见的雄激素敏感 LNCaP 和雄激素非依赖性 DU145 人细胞系中前列腺癌中 PCs 的功能冗余。我们使用基于慢病毒的 shRNA 传递系统,研究了内源性 PCs furin、PACE4 和 PC7 在这两种细胞系中的敲低细胞系的体外和体内细胞增殖特性。在这三种 PCs 中,只有 PACE4 是维持高增殖状态所必需的,这是通过体外 XTT 增殖测定和体内裸鼠肿瘤异种移植来确定的。在这两种细胞系中,furin 的敲低对细胞增殖或肿瘤异种移植生长均无影响。矛盾的是,PC7 的敲低降低了体外细胞增殖,但体内无影响。由于 PCs 作用于分泌途径,我们表明,来自 PACE4 敲低细胞的条件培养基在体外具有非常差的细胞生长刺激作用。PACE4 敲低肿瘤的免疫组织化学显示 Ki67 减少和 p27(KIP)水平升高(分别为增殖和细胞周期阻滞标志物)。有趣的是,我们确定仅在 PC7 敲低肿瘤中激活了表皮生长因子受体信号通路,这为 PC7 沉默在前列腺癌细胞系中的矛盾作用提供了一些解释。我们得出结论,PACE4 在维持前列腺癌的增殖和肿瘤进展中具有独特的作用,这使 PACE4 成为该疾病的一个相关治疗靶点。