Heilongjiang Key Laboratory of Scientific Research in Urology and Department of Urology, the Fourth Hospital of Harbin Medical University, Harbin, Heilongjiang Province, 150081, China.
Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine and Science, Rochester, MN 55905, USA.
Theranostics. 2019 May 24;9(12):3459-3475. doi: 10.7150/thno.33292. eCollection 2019.
The overall success rate of prostate cancer (PCa) diagnosis and therapy has been improved over the years. However, genomic and phenotypic heterogeneity remains a major challenge for effective detection and treatment of PCa. Efforts to better classify PCa into functional subtypes and elucidate the molecular mechanisms underlying prostate tumorigenesis and therapy resistance are warranted for further improvement of PCa outcomes. We generated ;cTg; ( double mutant) mice by crossbreeding ;cTg males with conditional () females. By using Hematoxylin and Eosin (H&E) staining, SMA and Masson's Trichrome staining, we investigated the effect of PTEN haploinsufficiency in combination with Runx2 overexpression on prostate tumorigenesis. Moreover, we employed immunohistochemistry (IHC) to stain Ki67 for cell proliferation, cleaved caspase 3 for apoptosis and AKT phosphorylation for signaling pathway in prostate tissues. Chromatin immunoprecipitation coupled quantitative PCR (ChIP-qPCR), reverse transcription coupled quantitative PCR (RT-qPCR), western blot (WB) analyses and immunofluorescence (IF) were conducted to determine the underlying mechanism by which RUNX2 regulates CXCR7 and AKT phosphorylation in PCa cells. We demonstrated that mice with prostate-specific heterozygous deletion and overexpression developed high-grade prostatic intraepithelial neoplasia (HGPIN) and cancerous lesions at age younger than one year, with concomitant high level expression of Akt phosphorylation and the chemokine receptor Cxcr7 in malignant glands. RUNX2 overexpression induced CXCR7 transcription and membrane location and AKT phosphorylation in PTEN-deficient human PCa cell lines. Increased expression of RUNX2 also promoted growth of PCa cells and this effect was largely mediated by CXCR7. CXCR7 expression also positively correlated with AKT phosphorylation in PCa patient specimens. Our results reveal a previously unidentified cooperative role of RUNX2 overexpression and PTEN haploinsufficiency in prostate tumorigenesis, suggesting that the defined RUNX2-CXCR7-AKT axis can be a viable target for effective treatment of PCa.
近年来，前列腺癌（PCa）的诊断和治疗整体成功率有所提高。然而，基因组和表型异质性仍然是有效检测和治疗 PCa 的主要挑战。为了进一步改善 PCa 的预后，有必要更好地将 PCa 分类为功能亚型，并阐明前列腺肿瘤发生和治疗耐药的分子机制。我们通过将雄性；cTg；与条件性（）雌性杂交，生成了；cTg；（双突变）小鼠。通过使用苏木精和伊红（H&E）染色、SMA 和 Masson's Trichrome 染色，我们研究了 PTEN 杂合不足与 Runx2 过表达相结合对前列腺肿瘤发生的影响。此外，我们采用免疫组织化学（IHC）染色 Ki67 以检测细胞增殖，cleaved caspase 3 以检测细胞凋亡，AKT 磷酸化以检测前列腺组织中的信号通路。染色质免疫沉淀结合定量 PCR（ChIP-qPCR）、逆转录定量 PCR（RT-qPCR）、western blot（WB）分析和免疫荧光（IF）用于确定 RUNX2 调节 PCa 细胞中 CXCR7 和 AKT 磷酸化的潜在机制。我们证明，前列腺特异性杂合缺失和过表达的小鼠在不到一年的年龄就发展出高级别前列腺上皮内瘤变（HGPIN）和癌性病变，同时恶性腺体中 AKT 磷酸化和趋化因子受体 Cxcr7 的表达水平升高。RUNX2 过表达诱导 PTEN 缺陷的人 PCa 细胞系中 CXCR7 的转录和膜定位以及 AKT 磷酸化。RUNX2 的过表达还促进了 PCa 细胞的生长，这种作用在很大程度上是由 CXCR7 介导的。CXCR7 的表达也与 PCa 患者标本中的 AKT 磷酸化呈正相关。我们的结果揭示了 RUNX2 过表达和 PTEN 杂合不足在前列腺肿瘤发生中的先前未被识别的协同作用，表明定义明确的 RUNX2-CXCR7-AKT 轴可以成为 PCa 有效治疗的可行靶点。
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