1. Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA;
J Cancer. 2013;4(4):296-303. doi: 10.7150/jca.5788. Epub 2013 Mar 20.
Human endometase/matrilysin-2/matrix metalloproteinase-26 (MMP-26) is an endopeptidase mostly produced by human carcinoma cells. While MMPs are thought to regulate the dynamics of extracellular matrix turnover, new evidence shows that these enzymes may play a critical regulatory role in inflammation. To investigate the role of MMP-26 in inflammation, three different variants of androgen repressed human prostate cancer (ARCaP) cells were investigated in the study: parental, MMP-26 sense cDNA-transfected, and MMP-26 antisense cDNA-transfected ARCaP cells. Protein lysates and RNA from control and genetically modified cells were analyzed by Western blotting and real-time reverse transcription polymerase chain reaction on arrays of genes critical to the inflammatory response. In comparison to parental controls, up-regulation of MMP-26 expression in MMP-26 sense cDNA-transfected cells resulted in a decrease in inflammatory genes expression. Conversely, inflammatory genes were up-regulated in MMP-26 antisense cDNA-transfected cells. Therefore, modulation of MMP-26 levels significantly affects the expression of inflammatory genes, suggesting an anti-inflammatory role of MMP-26. To determine a possible mechanism of action, further analysis, at both transcript and protein levels, revealed a dramatic down-regulation of interleukin-10 receptor B (IL10RB) in MMP-26 antisense cDNA-transfected cells. The low level of IL10RB was inversely correlated with matrix metalloproteinase-9 (MMP-9) expression. Collectively, our data suggest that the deficiency of MMP-26 may promote inflammation via inhibition of IL10RB-mediated signaling. These results propose a novel anti-inflammation function of MMP-26 and could provide novel molecular insight of therapeutic targeting.
人子宫内膜酶/基质金属蛋白酶-2(MMP-26)是一种主要由人癌细胞产生的内肽酶。虽然 MMP 被认为调节细胞外基质周转的动态平衡,但新的证据表明,这些酶可能在炎症中发挥关键的调节作用。为了研究 MMP-26 在炎症中的作用,研究中调查了三种不同的雄激素抑制人前列腺癌细胞(ARCaP)变体:亲本、MMP-26 感 cDNA 转染和 MMP-26 反义 cDNA 转染 ARCaP 细胞。通过 Western 印迹和实时逆转录聚合酶链反应对对照和遗传修饰细胞的蛋白裂解物和 RNA 进行分析,分析了对炎症反应至关重要的基因阵列。与亲本对照相比,MMP-26 感 cDNA 转染细胞中 MMP-26 表达的上调导致炎症基因表达的下调。相反,MMP-26 反义 cDNA 转染细胞中炎症基因上调。因此,MMP-26 水平的调节显著影响炎症基因的表达,表明 MMP-26 具有抗炎作用。为了确定可能的作用机制,在转录和蛋白水平上的进一步分析表明,MMP-26 反义 cDNA 转染细胞中白细胞介素-10 受体 B(IL10RB)的表达显著下调。IL10RB 的低水平与基质金属蛋白酶-9(MMP-9)的表达呈负相关。总的来说,我们的数据表明 MMP-26 的缺乏可能通过抑制 IL10RB 介导的信号转导促进炎症。这些结果提出了 MMP-26 的一种新的抗炎功能,并为治疗靶点提供了新的分子见解。