Tumor necrosis factor-alpha (TNFα) is an inflammatory cytokine that regulates inflammation and tumor progression in non-small cell lung cancer (NSCLC). The higher levels of TNF α are known to induce expression of several genes such as TNFα-induced protein 2 (TNFAIP2) with a largely unknown role in NSCLC. We provide the preliminary evidence for the role of TNFAIP2 in NSCLC progression and its epigenetic regulation mediated by microRNA, miR-145-5p. The expression of TNFAIP2 was confirmed using quantitative real-time PCR, immunohistochemistry, and Western blot in NSCLC tissue and paired adjacent normal tissue. All in vitro assays were undertaken in A549 and H23 cells and chemoresistance assays were undertaken in A549/Cisplatin (DDP) and H23/DDP cell types. TNFAIP2 silencing was undertaken using lipofectamine transfection of specific siRNA. Cells were co-transfected with miR-145-5p, and TNFAIP2–3′ untranslated region (UTR) or TNFAIP2 with mutated 3′UTR using the luciferase vector pGL. Cell viability, transwell migration, and invasion were assessed. The role of caspase 3 proteins in cell viability was ascertained using Western blot. The tumor tissues (and cisplatin-resistant cell lines A549/DDP and H23/DDP) expressed significantly higher levels of TNAIP2 mRNA and protein. Silencing of TNFAIP2 resulted in reduced cell viability, reduced invasion, and migration in vitro. Silencing of TNFAIP2 in A549/DDP and H23/DDP had higher expression of TNFAIP2, reduced cell viability, and increased induction of caspase 3. MiR-145-5p binds to the 3′UTR of TNFAIP2. Overexpression of MiR-145-5p reduced expression of TNFAIP2, decreased cell viability, reduced cell migration and invasion, and significantly reduced expression of caspase 3 protein. TNFAIP2 mediates tumorigenesis in NSCLC through, not completely known pathways. miR-145-5p negatively regulates TNFAIP2 expression. miR-145-5p-mediated therapies may be explored in NSCLC.

中文翻译：

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TNFAIP2促进非小细胞肺癌细胞并被miR-145-5p靶向。

肿瘤坏死因子-α（TNFα）是一种炎症性细胞因子，可调节非小细胞肺癌（NSCLC）中的炎症和肿瘤进展。已知较高水平的TNFα诱导几种基因的表达，例如TNFα诱导的蛋白2（TNFAIP2），其在NSCLC中的作用尚不清楚。我们提供了TNFAIP2在NSCLC进展中的作用及其由微小RNA miR-145-5p介导的表观遗传调控的初步证据。使用定量实时PCR，免疫组织化学和Western印迹法在NSCLC组织和配对的正常组织中证实了TNFAIP2的表达。全部体外在A549和H23细胞中进行测定，在A549 /顺铂（DDP）和H23 / DDP细胞类型中进行化学耐药性测定。使用脂质体转染特定siRNA进行TNFAIP2沉默。使用荧光素酶载体pGL ，将细胞与miR-145-5p和TNFAIP2-3 '非翻译区（UTR）或TNFAIP2与3'UTR突变共转染。评估细胞活力，跨孔迁移和入侵。使用蛋白质印迹法确定了胱天蛋白酶3蛋白在细胞活力中的作用。肿瘤组织（和顺铂耐药细胞系A549 / DDP和H23 / DDP）表达的TNAIP2 mRNA和蛋白水平明显较高。TNFAIP2沉默导致细胞活力降低，侵袭和体外迁移减少。在A549 TNFAIP2的沉默/ DDP和H23 / DDP有TNFAIP2的更高表达，降低细胞活力，和caspase 3 MIR-145-5p结合的诱导增加至的3'UTR TNFAIP2。MiR-145-5p的过表达降低了TNFAIP2的表达，降低了细胞活力，降低了细胞迁移和侵袭，并显着降低了caspase 3蛋白的表达。TNFAIP2通过未知途径介导NSCLC的肿瘤发生。miR-145-5p负调节TNFAIP2表达。miR-145-5p介导的疗法可在NSCLC中进行探索。