Hill K S, Erdogan E, Khoor A, Walsh M P, Leitges M, Murray N R, Fields A P
Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
Department of Pathology, Mayo Clinic, Jacksonville, FL, USA.
Oncogene. 2014 Apr 17;33(16):2134-44. doi: 10.1038/onc.2013.147. Epub 2013 Apr 22.
Protein kinase C alpha (PKCα) can activate both pro- and anti-tumorigenic signaling depending upon cellular context. Here, we investigated the role of PKCα in lung tumorigenesis in vivo. Gene expression data sets revealed that primary human non-small lung cancers (NSCLC) express significantly decreased PKCα levels, indicating that loss of PKCα expression is a recurrent event in NSCLC. We evaluated the functional relevance of PKCα loss during lung tumorigenesis in three murine lung adenocarcinoma models (LSL-Kras, LA2-Kras and urethane exposure). Genetic deletion of PKCα resulted in a significant increase in lung tumor number, size, burden and grade, bypass of oncogene-induced senescence, progression from adenoma to carcinoma and a significant decrease in survival in vivo. The tumor promoting effect of PKCα loss was reflected in enhanced Kras-mediated expansion of bronchio-alveolar stem cells (BASCs), putative tumor-initiating cells, both in vitro and in vivo. LSL-Kras/Prkca(-/-) mice exhibited a decrease in phospho-p38 MAPK in BASCs in vitro and in tumors in vivo, and treatment of LSL-Kras BASCs with a p38 inhibitor resulted in increased colony size indistinguishable from that observed in LSL-Kras/Prkca(-/-) BASCs. In addition, LSL-Kras/Prkca(-/-) BASCs exhibited a modest but reproducible increase in TGFβ1 mRNA, and addition of exogenous TGFβ1 to LSL-Kras BASCs results in enhanced growth similar to untreated BASCs from LSL-Kras/Prkca(-/-) mice. Conversely, a TGFβR1 inhibitor reversed the effects of PKCα loss in LSL-Kras/Prkca(-/-) BASCs. Finally, we identified the inhibitors of DNA binding (Id) Id1-3 and the Wilm's Tumor 1 as potential downstream targets of PKCα-dependent tumor suppressor activity in vitro and in vivo. We conclude that PKCα suppresses tumor initiation and progression, at least in part, through a PKCα-p38MAPK-TGFβ signaling axis that regulates tumor cell proliferation and Kras-induced senescence. Our results provide the first direct evidence that PKCα exhibits tumor suppressor activity in the lung in vivo.
蛋白激酶Cα(PKCα)可根据细胞环境激活促肿瘤和抗肿瘤信号传导。在此,我们研究了PKCα在体内肺肿瘤发生中的作用。基因表达数据集显示,原发性人类非小细胞肺癌(NSCLC)中PKCα水平显著降低,表明PKCα表达缺失是NSCLC中反复出现的事件。我们在三种小鼠肺腺癌模型(LSL-Kras、LA2-Kras和氨基甲酸乙酯暴露)中评估了PKCα缺失在肺肿瘤发生过程中的功能相关性。PKCα的基因缺失导致肺肿瘤数量、大小、负担和分级显著增加,癌基因诱导的衰老被绕过,从腺瘤进展为癌,并且体内存活率显著降低。PKCα缺失的促肿瘤作用体现在体外和体内Kras介导的支气管肺泡干细胞(BASC)(假定的肿瘤起始细胞)扩增增强。LSL-Kras/Prkca(-/-)小鼠在体外BASC和体内肿瘤中的磷酸化p38 MAPK水平降低,用p38抑制剂处理LSL-Kras BASC导致集落大小增加,与LSL-Kras/Prkca(-/-) BASC中观察到的情况无法区分。此外,LSL-Kras/Prkca(-/-) BASC中TGFβ1 mRNA有适度但可重复的增加,向LSL-Kras BASC中添加外源性TGFβ1导致生长增强,类似于未处理的LSL-Kras/Prkca(-/-)小鼠的BASC。相反,TGFβR1抑制剂逆转了LSL-Kras/Prkca(-/-) BASC中PKCα缺失的影响。最后,我们确定DNA结合抑制剂(Id)Id1-3和威尔姆斯瘤1为体外和体内PKCα依赖性肿瘤抑制活性的潜在下游靶点。我们得出结论,PKCα至少部分通过调节肿瘤细胞增殖和Kras诱导的衰老的PKCα-p38MAPK-TGFβ信号轴抑制肿瘤起始和进展。我们的结果提供了首个直接证据,表明PKCα在体内肺中表现出肿瘤抑制活性。