Targeting cathepsin K diminishes prostate cancer establishment and growth in murine bone
Targeting cathepsin K diminishes prostate cancer establishment and growth in murine bone
复制标题
靶向组织蛋白酶 K 可减少前列腺癌在小鼠骨骼中的形成和生长
DOI：
10.1007/s00432-019-02950-y
复制
发表时间：
2019-08-01
影响因子：
3.6
通讯作者：
Zhang，Jian
中科院分区：
文献类型：
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作者：
Liang，Weiping;Wang，Fuhao;Zhang，Jian
背景前列腺癌（prostate cancer，PCa）的侵袭和转移过程是由多种蛋白酶参与的蛋白水解级联过程，包括基质金属蛋白酶（matrix metalloproteinases）、丝氨酸蛋白酶（serine proteases）和半胱氨酸蛋白酶（cathepsin K，CatK）。CatK主要由破骨细胞分泌，并特异性降解胶原蛋白I，导致骨破坏。前列腺癌和乳腺癌优先转移到骨。重要的是，与原发性肿瘤和正常前列腺组织相比，CatK表达水平在PCa骨转移部位更高。然而，CatK在PCa转移到骨中的潜在机制仍有待阐明。我们研究了CatK在PCa的建立和生长过程中的功能作用，在小鼠bone.MethodsCatK mRNA的表达进行了验证，通过免疫印迹在PCa LNCaP，C4 - 2B，和PC3细胞以及PCa组织中的蛋白表达。用ELISA法测定其蛋白质产量。比较了通过siRNA和CatK抑制剂的敲减对PCa细胞侵袭的影响。我们进一步研究了CatK抑制剂对条件培养基诱导的骨吸收的剂量依赖性作用。在建立动物模型时，将C4 - 2B细胞注射到SCID小鼠的胫骨中。在肿瘤细胞注射（肿瘤建立模型;协议I）或肿瘤细胞注射（肿瘤进展模型;协议II）后4周用载体或CatK抑制剂治疗8周的动物被应用于组织学和histomorphometric analysis.ResultsWe确认CatK在PCa LNCaP，C4 - 2B和PC3细胞以及PCa组织中的表达。此外，我们观察了选择性CatK抑制剂对PCa细胞侵袭的抑制作用。CatK抑制剂剂量依赖性地抑制PCA条件培养基诱导的骨吸收。在将C4 - 2B细胞注射到SCID小鼠的胫骨中时，选择性CatK抑制剂在方案I中显著防止肿瘤建立，并且在方案II中减少骨中的肿瘤生长。它还降低了两种动物模型中的血清PSA水平。CatK抑制剂与唑来膦酸（zoledronic acid，ZA）联合应用可增强其抑制作用。结论选择性CatK抑制剂可预防骨PCa的发生和发展，为晚期PCa的治疗提供了新的途径。
BackgroundThe processes of prostate cancer (PCa) invasion and metastasis are facilitated by proteolytic cascade involving multiple proteases, such as matrix metalloproteinases, serine proteases and cysteine proteases including cathepsin K (CatK). CatK is predominantly secreted by osteoclasts and specifically degrades collagen I leading to bone destruction. PCa and breast cancer preferentially metastasize to the bone. Importantly, CatK expression level is greater in PCa bone metastatic sites compared to primary tumor and normal prostate tissues. However, the underlying mechanism of CatK during PCa metastases into the bone remains to be elucidated. We investigated the functional role of CatK during the PCa establishment and growth process in the murine bone.MethodsCatK mRNA expression was validated by RT-PCR, protein expression by immunoblotting in PCa LNCaP, C4-2B, and PC3 cells as well as in PCa tissues. Its protein production was measured using ELISA assay. The effect of both knockdowns via siRNA and CatK inhibitor was compared in regard to PCa cell invasion. We further studied the dose-dependent CatK inhibitor effect on conditioned media-induced bone resorption. In setting up an animal model, C4-2B cells were injected into the tibiae of SCID mice. The animals treated with either vehicle or CatK inhibitor for 8 weeks at the time of tumor cell injection (tumor establishment model; protocol I) or 4 weeks after tumor cell injection (tumor progression model; protocol II) were applied to histological and histomorphometric analyses.ResultsWe confirmed CatK expression in PCa LNCaP, C4-2B, and PC3 cells as well as in PCa tissues. Furthermore, we observed the inhibitory effects of a selective CatK inhibitor on PCa cell invasion. The CatK inhibitor dose-dependently inhibited PCa-conditioned media-induced bone resorption. Upon injection of C4-2B cells into the tibiae of SCID mice, the selective CatK inhibitor significantly prevented the tumor establishment in protocol I, and reduced the tumor growth in bone in protocol II. It also decreased serum PSA levels in both animal models. The inhibitory effects of the CatK inhibitor were enhanced in combination with zoledronic acid (ZA).ConclusionThe selective CatK inhibitor may prevent the establishment and progression of PCa in bone, thus making it a novel therapeutic approach for advanced PCa.