Kadasah Sultan F, Alqahtani Abdulaziz M S
Department of Biology, Faculty of Science, University of Bisha, P.O. Box 551, Bisha 61922, Saudi Arabia.
Int J Mol Sci. 2025 Jul 7;26(13):6532. doi: 10.3390/ijms26136532.
Lung adenocarcinoma is a prevalent, aggressive cancer with a poor prognosis due to early metastasis and resistance to treatment. LncRNA AFAP1-AS1 has been shown to be associated with the development of multiple carcinomas. This study investigates the functional role of AFAP1-AS1 in lung adenocarcinoma cell proliferation via miR-508-3p and ZWINT. Human lung adenocarcinoma A549 cells were transfected with siRNA constructs against AFAP1-AS1 (si-AFAP1-AS1) to silence its expression. Cell proliferation was evaluated via CCK-8 and colony-forming assays. Apoptosis was assessed using AO/EB staining, and invasion was determined via Transwell assay. The interaction between AFAP1-AS1, miR-508-3p, and ZWINT was confirmed via dual luciferase reporter assay and qRT-PCR analysis. Data were analysed using appropriate statistical tests. AFAP1-AS1 was significantly upregulated in lung adenocarcinoma cells compared to normal BEAS-2B cells. Silencing of AFAP1-AS1 resulted in a marked reduction in A549 cell proliferation and colony development, as observed in CCK-8 and colony formation assays. The AO/EB assay showed a significant increase in apoptosis (30 ± 4.4%) in si-AFAP1-AS1 transfected cells compared to control si-NC (3 ± 1.2%). In addition, knockdown of AFAP1-AS1 led to an upsurge of pro-apoptotic Bax and decline of anti-apoptotic Bcl-2 expression. The dual luciferase assay established the interaction between AFAP1-AS1 and miR-508-3p. Furthermore, ZWINT, identified as a target of miR-508-3p, was significantly upregulated in lung adenocarcinoma tissues. Overexpression of ZWINT rescued the inhibitory effects of AFAP1-AS1 silencing on cell proliferation, colony formation, and apoptosis, while also reversing the reduction in cell invasion. AFAP1-AS1 accelerates the development of lung adenocarcinoma by cell proliferation, apoptosis, and invasion via the miR-508-3p/ZWINT axis. Thus, targeting AFAP1-AS1 or its downstream regulatory axis could offer novel therapeutic approaches in lung adenocarcinoma treatment.
肺腺癌是一种常见的侵袭性癌症,由于早期转移和对治疗的抵抗性,其预后较差。长链非编码RNA AFAP1-AS1已被证明与多种癌症的发生发展有关。本研究通过miR-508-3p和ZWINT探讨AFAP1-AS1在肺腺癌细胞增殖中的功能作用。用针对AFAP1-AS1的小干扰RNA构建体(si-AFAP1-AS1)转染人肺腺癌A549细胞以沉默其表达。通过CCK-8和集落形成试验评估细胞增殖。使用AO/EB染色评估细胞凋亡,并通过Transwell试验测定细胞侵袭能力。通过双荧光素酶报告基因试验和qRT-PCR分析证实AFAP1-AS1、miR-508-3p和ZWINT之间的相互作用。使用适当的统计检验对数据进行分析。与正常BEAS-2B细胞相比,肺腺癌细胞中AFAP1-AS1显著上调。如CCK-8和集落形成试验所示,沉默AFAP1-AS1导致A549细胞增殖和集落形成明显减少。AO/EB试验显示,与对照小干扰RNA(si-NC)(3±1.2%)相比,转染si-AFAP1-AS1的细胞凋亡显著增加(30±4.4%)。此外,敲低AFAP1-AS1导致促凋亡蛋白Bax表达增加,抗凋亡蛋白Bcl-2表达下降。双荧光素酶试验证实了AFAP1-AS1与miR-508-3p之间的相互作用。此外,作为miR-508-3p靶点的ZWINT在肺腺癌组织中显著上调。ZWINT的过表达挽救了AFAP1-AS1沉默对细胞增殖、集落形成和凋亡的抑制作用,同时也逆转了细胞侵袭能力的降低。AFAP1-AS1通过miR-508-3p/ZWINT轴促进肺腺癌的发生发展,通过细胞增殖、凋亡和侵袭发挥作用。因此,靶向AFAP1-AS1或其下游调控轴可能为肺腺癌治疗提供新的治疗方法。