Horváth Zsolt, Kovalszky Ilona, Fullár Alexandra, Kiss Katalin, Schaff Zsuzsa, Iozzo Renato V, Baghy Kornélia
1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary.
2nd Department of Pathology, Semmelweis University, Budapest, Hungary.
Matrix Biol. 2014 Apr;35:194-205. doi: 10.1016/j.matbio.2013.11.004. Epub 2013 Dec 18.
Hepatocellular carcinoma represents one of the most-rapidly spreading cancers in the world. In the majority of cases, an inflammation-driven fibrosis or cirrhosis precedes the development of the tumor. During malignant transformation, the tumor microenvironment undergoes qualitative and quantitative changes that modulate the behavior of the malignant cells. A key constituent for the hepatic microenvironment is the small leucine-rich proteoglycan decorin, known to interfere with cellular events of tumorigenesis mainly by blocking various receptor tyrosine kinases (RTK) such as EGFR, Met, IGF-IR, PDGFR and VEGFR2. In this study, we characterized cell signaling events evoked by decorin deficiency in two experimental models of hepatocarcinogenesis using thioacetamide or diethyl nitrosamine as carcinogens. Genetic ablation of decorin led to enhanced tumor occurrence as compared to wild-type animals. These findings correlated with decreased levels of the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) and a concurrent elevation in retinoblastoma protein phosphorylation via cyclin dependent kinase 4. Decreased steady state p21(Waf1/Cip1) levels correlated with enhanced expression of transcription factor AP4, a known transcriptional repressor of p21(Waf1/Cip1), and enhanced c-Myc protein levels. In addition, translocation of β-catenin was a typical event in diethyl nitrosamine-evoked tumors. In parallel, decreased phosphorylation of both c-Myc and β-catenin was observed in Dcn(-/-) livers likely due to the hindered GSK3β-mediated targeting of these proteins to proteasomal degradation. We discovered that in a genetic background lacking decorin, four RTKs were constitutively activated (phosphorylated), including three known targets of decorin such as PDGFRα, EGFR, IGF-IR, and a novel RTK MSPR/RON. Our findings provide powerful genetic evidence for a crucial in vivo role of decorin during hepatocarcinogenesis as lack of decorin in the liver and hepatic stroma facilitates experimental carcinogenesis by providing an environment devoid of this potent pan-RTK inhibitor. Thus, our results support future utilization of decorin as an antitumor agent in liver cancer.
肝细胞癌是世界上传播速度最快的癌症之一。在大多数情况下,炎症驱动的纤维化或肝硬化先于肿瘤的发生。在恶性转化过程中,肿瘤微环境会发生定性和定量变化,从而调节恶性细胞的行为。肝微环境的一个关键成分是富含亮氨酸的小分子蛋白聚糖核心蛋白聚糖,已知它主要通过阻断各种受体酪氨酸激酶(RTK),如表皮生长因子受体(EGFR)、间质表皮转化因子(Met)、胰岛素样生长因子1型受体(IGF-IR)、血小板衍生生长因子受体(PDGFR)和血管内皮生长因子受体2(VEGFR2),来干扰肿瘤发生的细胞事件。在本研究中,我们使用硫代乙酰胺或二乙基亚硝胺作为致癌物,在两种肝癌发生的实验模型中,对核心蛋白聚糖缺乏引起的细胞信号事件进行了表征。与野生型动物相比,核心蛋白聚糖的基因缺失导致肿瘤发生率增加。这些发现与细胞周期蛋白依赖性激酶抑制剂p21(WAF1/CIP1)水平降低以及视网膜母细胞瘤蛋白通过细胞周期蛋白依赖性激酶4磷酸化同时升高相关。p21(Waf1/Cip1)稳态水平降低与转录因子AP4表达增强相关,AP4是已知的p21(Waf1/Cip1)转录抑制因子,同时c-Myc蛋白水平也增强。此外,β-连环蛋白的易位是二乙基亚硝胺诱发肿瘤中的典型事件。同时在Dcn(-/-)肝脏中观察到c-Myc和β-连环蛋白的磷酸化降低,这可能是由于糖原合成酶激酶3β(GSK3β)介导的这些蛋白靶向蛋白酶体降解受阻所致。我们发现,在缺乏核心蛋白聚糖的遗传背景下,四种RTK被组成性激活(磷酸化),包括核心蛋白聚糖的三个已知靶点,如PDGFRα、EGFR、IGF-IR,以及一种新的RTK MSPR/RON。我们的发现为核心蛋白聚糖在肝癌发生过程中的关键体内作用提供了有力的遗传学证据,因为肝脏和肝基质中缺乏核心蛋白聚糖,通过提供一个缺乏这种强效泛RTK抑制剂的环境,促进了实验性致癌作用。因此,我们的结果支持未来将核心蛋白聚糖用作肝癌的抗肿瘤药物。