A Strong Decrease in TIMP3 Expression Mediated by the Presence of miR-17 and 20a Enables Extracellular Matrix Remodeling in the NSCLC Lesion Surroundings
A Strong Decrease in TIMP3 Expression Mediated by the Presence of miR-17 and 20a Enables Extracellular Matrix Remodeling in the NSCLC Lesion Surroundings
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DOI：
10.3389/fonc.2019.01372
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发表时间：
2019-12-13
影响因子：
4.7
通讯作者：
Brzeziariska-Lasota, Ewa
中科院分区：
文献类型：
--
作者：
Czarnecka, Karolina H.;Szmyd, Bartosz;Brzeziariska-Lasota, Ewa
背景资料：肺癌是世界范围内最常见的死亡原因之一，具有相对高的病死率和约18%的平均5年生存率。肿瘤的标志之一是细胞外基质（ECM）重塑，这对转移至关重要。这一过程可以通过靶向与ECM分解和转移过程相关的金属蛋白酶（MMP）或阻断金属蛋白酶组织抑制剂（TIMP）的作用的miR来调节。寻找早期生物标志物对于检测非小细胞肺癌（NSCLC）并区分其亚型（腺癌（AC）和鳞状细胞癌（SCC））至关重要，从而实现靶向化疗。方法：通过TCGA数据分析筛选出靶向MMP 2和TIMP 3的miR-17和miR-20 a，并利用miRTarBase和文献进一步验证。研究组包括47例原发性NSCLC（AC和SCC亚型）患者。从肿瘤和不含癌细胞的正常外观邻近组织（NLNT）中分离RNA。在入院时和手术后5-7天从外周血外来体提取miR。使用TaqMan探针在qPCR中评估基因和miR表达。结果：MMP 2在NLNT中的表达水平与癌症相似。TIMP 3在癌组织和NLNT中的表达均降低，且在癌组织中的表达显著降低。TIMP 3在NLNT和SCC亚型中的下调与miR-20 a呈负相关。与AC相比，SCC患者的术前miR-17表达显著更高。miR-17作为AC亚型分类器的受试者工作特征（ROC）分析显示，在最佳截止点处的特异性为90%，灵敏度为48%，ROC曲线下面积（AUC）为0.71（95%CI：0.55-0.87）。在NSCLC亚型中：在SCC组中观察到NLNT的包年（PY）和TIMP 3表达之间存在强负相关性。总结：在NLNT中观察到的TIMP 3沉默及其与miR-20 a（来自血清外泌体）的术前表达的负相关性表明miR可以在距离病变中心一定距离处影响ECM重塑。手术前获得的血清中的miR表达模式在AC和SCC亚型之间显著不同。此外，NLNT（SCC组）中TIMP 3表达降低与PY一生中吸烟量呈负相关。
Background: Lung cancer is one of the most common causes of death worldwide with a relatively high fatality rate and a mean 5-years survival of about 18%. One of the hallmarks of cancer is the extracellular matrix (ECM) remodeling, which is crucial for metastasis. This process may be regulated by miRs targeting metalloproteinases (MMPs) associated with the ECM breakdown and metastatic process or blocking the action of tissue inhibitors of metalloproteinases (TIMPs). Search for early biomarkers is essential in detecting non-small cell lung cancer (NSCLC) and distinguishing its subtypes: Adenocarcinoma (AC) from Squamous Cell Carcinoma (SCC), enabling targeted chemotherapy. Methods: MiR-17 and miR-20a targeting MMP2 and TIMP3 were selected by TCGA data analysis with further validation using miRTarBase and literature. The study group comprised 47 patients with primary NSCLC (AC and SCC subtypes). RNA was isolated from the tumor and normal-looking neighboring tissue (NLNT) free of cancer cells. MiRs from peripheral blood exosomes were extracted on admission and 5-7 days after surgery. Gene and miRs expression were assessed in qPCR using TaqMan probes. Results: The MMP2 has been expressed on a similar level in NLNT, as in cancer. While, TIMP3 expression was decreased both in cancer tissue and NLNT, with significantly lower expression in cancer. TIMP3 downregulation in NLNT and in SCC subtype correlated negatively with miR-20a. The preoperative miR-17 expression was significantly higher among patients with SCC compared to AC. Receiver operating characteristic (ROC) analysis of miR-17 as AC subtype classifier revealed 90% specificity and 48% sensitivity in optimal cut-off point with area under ROC curve (AUC): 0.71 (95%CI: 0.55-0.87). Within NSCLC subtypes: a strong negative correlation between pack-years (PY) and TIMP3 expression was observed for NLNT in the SCC group. Conclusion: The TIMP3 silencing observed in the NLNT and its negative correlation with presurgical expression of miR-20a (from serum exosomes), suggest that miRs can influence ECM remodeling at a distance from the center of the lesion. The miRs expression pattern in serum obtained before surgery significantly differs between AC and SCC subtypes. Moreover, decreased TIMP3 expression in NLNT (in SCC group) negatively correlates with the amount of tobacco smoked in a lifetime in PY.