RESUMO
BACKGROUND: Long intergenic non-coding RNA 326 (LINC00326) modulates hepatocarcinogenic lipid metabolism. However, the ability of LINC00326 to modulate the highly aggressive non-small cell lung carcinoma (NSCLC) is unknown. Here, LINC00326 in NSCLC was investigated, together with its effects on tumor malignancy and the underlying mechanisms of action. METHODS: LINC00326 levels in tumor tissues and cell lines were measured by Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and RNA fluorescence in situ hybridization (FISH). Proliferation and apoptosis were assessed in cell lines by Cell Counting Kit-8 (CCK-8), EdU staining assays and flow cytometry, respectively, and tumor growth was measured in mouse models. Possible microRNA targets of LINC00326 were predicted by bioinformatics and verified by RNA pull-down and immunoprecipitation and luciferase reporter assays. Western blotting was used to evaluate the expression of Wnt/ß-catenin-associated proteins. RESULTS: LINC00326 was downregulated in tumor tissues and cell lines. Knockdown of LINC00326 stimulated NSCLC cell proliferation and suppressed apoptosis in vitro, as well as enhancing xenograft tumor growth. LINC00326 sponged miR-657, and dickkopf WNT signaling pathway inhibitor 2 (DKK2) was found to be directly targeted by miR-657, with LINC00326 positively regulating its expression through sponging miR-657. The actions of LINC00326 knockdown on proliferation and apoptosis were reversed by stimulation of the miR-657/DKK2 axis. Furthermore, overexpression of miR-657 mitigated DKK2 inhibition on Wnt/ß-catenin signaling. CONCLUSIONS: LINC00326/miR-657/DKK2 axis signaling blocked tumor-associated functions in NSCLC cells through the targeting Wnt/ß-catenin pathway. This suggests that this pathway could be a target for NSCLC treatment.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , MicroRNAs , RNA Longo não Codificante , Animais , Camundongos , Humanos , Carcinoma Pulmonar de Células não Pequenas/genética , Via de Sinalização Wnt/genética , beta Catenina/genética , Regulação para Cima , Hibridização in Situ Fluorescente , Linhagem Celular Tumoral , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias Pulmonares/genética , Proliferação de Células/genética , RNA Longo não Codificante/genética , Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica , Apoptose/genéticaRESUMO
Background: The purpose of our study was to evaluate whether the methylation status of the miR-657 promoter region could be used as a biomarker for diagnosis of hepatocellular carcinoma (HCC), so as to find alternative biomarkers of early HCC detection. Methods: Cancerous and paired adjacent noncancerous tissues were collected from 160 patients who had been diagnosed with HCC by histopathology and received surgery. The methylation status of the miR-657 promoter region was measured using a MassARRAY Analyzer 4. Receiver operator characteristic (ROC) curve analysis was used to assess the effectiveness of miR-657 promoter region methylation status as a biomarker for diagnosis of HCC. Results: The mean methylation level of the miR-657 promoter region was significantly lower in cancerous tissues than in normal tissues of HCC patients (48.91%:67.04%, P<0.0001). ROC curve analysis revealed that the mean methylation level of the miR-657 promoter region could distinguish cancerous tissues from paired normal tissues of HCC patients (area under the curve: 0.847, P<0.001). Using 59.50% as the optimal cut-off, the sensitivity was 95.50% and the specificity was 70.01%. Conclusions: Methylation levels of the miR-657 promoter region were decreased in HCC patients and could be used as alternative and supplementary biomarkers for diagnosis of HCC.
RESUMO
Though Spatholobus suberectus Dunn (SSD) has been reported to have anti-virus, anti-osteoclastogenesis, and anti-inflammation activities, its underlying anti-cancer mechanism has never been elucidated in association with the role of miR-657 in endoplasmic reticulum (ER) stress-related apoptosis to date. SSD treatment exerted cytotoxicity in U266 and U937 cells in a dose-dependent manner. Also, apoptosis-related proteins such as PARP, procaspase-3, and Bax were regulated by SSD treatment. Furthermore, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay revealed that a number of apoptotic bodies were increased by SSD. Interestingly, the ER stress-related proteins such as p-ATF2 and CHOP were elevated by SSD. Interestingly, reactive oxygen species (ROS) generation and cytotoxicity by SSD treatment were significantly reduced by N-Acetyl-L-cysteine (NAC). Among the microRNAs (miRNAs) regulated by SSD treatment, miR-657 was most significantly reduced by SSD treatment. However, an miR-657 mimic reversed SSD-induced apoptosis by the attenuation of the expression of p-ATF2, CHOP, and PARP cleavage. Overall, these findings provide scientific evidence that miR657 is an onco-miRNA targeting the ER stress signal pathway and SSD induces apoptosis via the inhibition of miR-657, ROS, and the activation of p-ATF2 and CHOP as a potent anti-cancer agent for myeloid-originated hematological cancer.