School of Basic Medical Science, Southwest Medical University, Luzhou 646000, China.
Department of Respirology and Critical Care Medicine, Chengdu Seventh People's Hospital, Chengdu 610041, China.
Dis Markers. 2022 Aug 17;2022:4842454. doi: 10.1155/2022/4842454. eCollection 2022.
MicroRNA- (miR-) 657 has been shown to regulate immunological and inflammatory activity, and it has also been defined to be dysregulated in both non-small-cell lung cancer (NSCLC) and hepatocellular carcinoma. The mechanistic role whereby miR-657 influences NSCLC progression, however, has yet to be clarified.
miR-657 and SRCIN1 expression levels were assessed via qPCR in the cell lines and tissues of NSCLC. Besides, correlations between the levels of miR-657 and NSCLC patient pathological characteristics were examined, and the Kaplan-Meier approach was employed for the evaluation of the prognostic utility of miR-657 in these patients. Moreover, the Pearson correlation analyses and dual-luciferase reporter assessments were used for detecting interactive relationships between miR-657 and SRCIN1. In addition, CCK-8, EdU, and Transwell assessments were employed for the appraisal of the ability of miR-657/SRCIN1 to regulate NSCLC cell proliferation and invasion. Western blotting was employed for the assessment of the levels of NSCLC cell proteins associated with the epithelial-mesenchymal transition (EMT) that were influenced by miR-657. The nude mice xenograft tumor model is established to observe the effect of miR-657 on NSCLC growth .
NSCLC patient tissues and cell lines exhibited upregulated miR-657 expression that was closely related to tumor differentiation, lymphoid metastasis, and TNM stage. High levels of miR-657 were predictive of a poorer NSCLC patient prognosis, and overexpressing miR-657 resulted in the more rapid growth of NCI-H1650 and A549 cells, with a concomitant increase in their invasion. In addition, miR-657 overexpression raised the levels of Slug, N-cadherin, and Vimentin in these two cell lines while promoting E-cadherin downregulation. Dual-luciferase reporter assays confirmed that miR-657 was capable of binding to the SRCIN1 gene, and SRCIN1 expression levels were negatively associated with those of miR-657, indicating that it acts as a negative regulator of this gene. Knocking down SRCIN1 was capable to reverse the influences of miR-657 inhibitor treatment on NSCLC cell behavior. Finally, studies showed that miR-657 promoted NSCLC cell growth.
The obtained findings illuminate that miR-657 can promote the growth of tumors and the induction of the EMT in NSCLC cells by targeting SRCIN1 expression and modulating Slug pathway activation, highlighting this pathway as a promising therapeutic target in cases suffering from NSCLC.
MicroRNA-（miR-）657 已被证明可调节免疫和炎症活性，并且在非小细胞肺癌（NSCLC）和肝细胞癌中也发现其失调。然而，miR-657 影响 NSCLC 进展的机制作用尚不清楚。
通过 qPCR 检测 NSCLC 细胞系和组织中的 miR-657 和 SRCIN1 表达水平。此外，还检查了 miR-657 水平与 NSCLC 患者病理特征之间的相关性，并采用 Kaplan-Meier 方法评估 miR-657 在这些患者中的预后效用。此外，还进行了 Pearson 相关性分析和双荧光素酶报告基因评估，以检测 miR-657 和 SRCIN1 之间的相互作用关系。另外，还采用 CCK-8、EdU 和 Transwell 评估来评估 miR-657/SRCIN1 调节 NSCLC 细胞增殖和侵袭的能力。采用 Western blot 检测 miR-657 影响的与上皮-间充质转化（EMT）相关的 NSCLC 细胞蛋白水平。建立裸鼠异种移植肿瘤模型观察 miR-657 对 NSCLC 生长的影响。
NSCLC 患者组织和细胞系中 miR-657 表达上调，与肿瘤分化、淋巴转移和 TNM 分期密切相关。高水平的 miR-657 预示着 NSCLC 患者预后较差，过表达 miR-657 导致 NCI-H1650 和 A549 细胞生长更快，侵袭能力增强。此外，miR-657 过表达增加了这两种细胞系中 Slug、N-钙粘蛋白和波形蛋白的水平，同时促进了 E-钙粘蛋白的下调。双荧光素酶报告基因实验证实 miR-657 能够与 SRCIN1 基因结合，并且 SRCIN1 表达水平与 miR-657 呈负相关，表明其作为该基因的负调节剂。敲低 SRCIN1 能够逆转 miR-657 抑制剂处理对 NSCLC 细胞行为的影响。最后，研究表明 miR-657 可促进 NSCLC 细胞生长。
研究结果表明，miR-657 通过靶向 SRCIN1 表达和调节 Slug 通路激活，促进 NSCLC 细胞肿瘤生长和 EMT 的诱导，凸显了该通路作为治疗 NSCLC 的潜在治疗靶点。
