Tao Min, Liu Lu, Shen Meng, Zhi Qiaoming, Gong Fei-Ran, Zhou Binhua P, Wu Yadi, Liu Haiyan, Chen Kai, Shen Bairong, Wu Meng-Yao, Shou Liu-Mei, Li Wei
a Department of Oncology , the First Affiliated Hospital of Soochow University , Suzhou , China.
b PREMED Key Laboratory for Precision Medicine, Soochow University , Suzhou , China.
Cell Cycle. 2016;15(3):381-93. doi: 10.1080/15384101.2015.1127468.
Previous studies have indicated that inflammatory stimulation represses protein phosphatase 2A (PP2A), a well-known tumor suppressor. However, whether PP2A repression participates in pancreatic cancer progression has not been verified. We used lipopolysaccharide (LPS) and macrophage-conditioned medium (MCM) to establish in vitro inflammation models, and investigated whether inflammatory stimuli affect pancreatic cancer cell growth and invasion PP2A catalytic subunit (PP2Ac)-dependently. Via nude mouse models of orthotopic tumor xenografts and dibutyltin dichloride (DBTC)-induced chronic pancreatitis, we evaluated the effect of an inflammatory microenvironment on PP2Ac expression in vivo. We cloned the PP2Acα and PP2Acβ isoform promoters to investigate the PP2Ac transcriptional regulation mechanisms. MCM accelerated pancreatic cancer cell growth; MCM and LPS promoted cell invasion. DBTC promoted xenograft growth and metastasis, induced tumor-associated macrophage infiltration, promoted angiogenesis, activated the nuclear factor-κB (NF-κB) pathway, and repressed PP2Ac expression. In vitro, LPS and MCM downregulated PP2Ac mRNA and protein. PP2Acα overexpression attenuated JNK, ERK, PKC, and IKK phosphorylation, and impaired LPS/MCM-stimulated cell invasion and MCM-promoted cell growth. LPS and MCM activated the NF-κB pathway in vitro. LPS and MCM induced IKK and IκB phosphorylation, leading to p65/RelA nuclear translocation and transcriptional activation. Overexpression of the dominant negative forms of IKKα attenuated LPS and MCM downregulation of PP2Ac, suggesting inflammatory stimuli repress PP2Ac expression NF-κB pathway-dependently. Luciferase reporter gene assay verified that LPS and MCM downregulated PP2Ac transcription through an NF-κB-dependent pathway. Our study presents a new mechanism in inflammation-driven cancer progression through NF-κB pathway-dependent PP2Ac repression.
先前的研究表明,炎症刺激会抑制蛋白磷酸酶2A(PP2A),这是一种著名的肿瘤抑制因子。然而,PP2A抑制是否参与胰腺癌进展尚未得到证实。我们使用脂多糖(LPS)和巨噬细胞条件培养基(MCM)建立体外炎症模型,并研究炎症刺激是否通过PP2A催化亚基(PP2Ac)依赖性方式影响胰腺癌细胞的生长和侵袭。通过原位肿瘤异种移植裸鼠模型和二丁基二氯化锡(DBTC)诱导的慢性胰腺炎,我们评估了炎症微环境对体内PP2Ac表达的影响。我们克隆了PP2Acα和PP2Acβ亚型启动子以研究PP2Ac的转录调控机制。MCM加速了胰腺癌细胞的生长;MCM和LPS促进了细胞侵袭。DBTC促进了异种移植瘤的生长和转移,诱导了肿瘤相关巨噬细胞浸润,促进了血管生成,激活了核因子κB(NF-κB)通路,并抑制了PP2Ac的表达。在体外,LPS和MCM下调了PP2Ac的mRNA和蛋白水平。PP2Acα的过表达减弱了JNK、ERK、PKC和IKK的磷酸化,并损害了LPS/MCM刺激的细胞侵袭和MCM促进的细胞生长。LPS和MCM在体外激活了NF-κB通路。LPS和MCM诱导了IKK和IκB的磷酸化,导致p65/RelA核转位和转录激活。IKKα显性负性形式的过表达减弱了LPS和MCM对PP2Ac的下调,表明炎症刺激通过NF-κB通路依赖性方式抑制PP2Ac的表达。荧光素酶报告基因检测证实,LPS和MCM通过NF-κB依赖性途径下调PP2Ac的转录。我们的研究揭示了炎症驱动癌症进展过程中通过NF-κB通路依赖性PP2Ac抑制的新机制。