Zhu Wangyu, Yu Yun, Ye Yuxin, Tu Xinyue, Zhang Yan, Wu Tao, Ni Lianli, Huang Xiangjie, Wang Yumin, Cui Ri
Cellular and Molecular Biology Laboratory, Affiliated Zhoushan Hospital of Wenzhou Medical University, Zhoushan, Zhejiang 316020, China; Cancer and Anticancer Drug Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
Cancer and Anticancer Drug Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
Transl Oncol. 2023 Nov;37:101755. doi: 10.1016/j.tranon.2023.101755. Epub 2023 Aug 16.
Our recent study found that QKI-5 regulated miRNA, miR-196b-5p, promotes non-small cell lung cancer (NSCLC) progression by directly targeting GATA6, TSPAN12 and FAS. However, the biological functions of miR-196b-5p in NSCLC progression and metastasis still remain elusive.
Cell proliferation, migration, colony formation, cell cycle assays were used to investigate cellular phenotypic changes. Quantitative real-time PCR (qRT-PCR) and western blot analyses were used to measure expressions of relative gene and protein. Interaction between QKI-5 and miR-196b-5p was determined by RNA immunoprecipitation (RIP) assay. Luciferase reporter assay was used to determine direct binding between miR-196b-5p and NFKBIA 3'-UTR. ELISA assay was used to measure secreted IL6 proteins. Mice xenograft model was used to assess the functions of NFKBIA on in vivo tumor growth.
We demonstrated that the miR-196b-5p facilitates lung cancer cell proliferation, migration, colony formation, and cell cycle by directly targeting NFKBIA, a negative regulator of NFκB signaling. Knocking down NFKBIA increases IL6 mediated phosphorylation of signal transducer and activator of transcription 3 (STAT3) to promote lung cancer cell growth by activating NFκB signaling. The expression of NFKBIA was significantly downregulated in NSCLC tissue samples, and was negatively correlated with the expression miR-196b-5p. In addition, we found that downregulated QKI-5 expression was associated with the elevated miR-224 expression in NSCLC.
Our findings indicated that the miR-224/QKI-5/miR-196b-5p/NFKBIA signaling pathway might play important functions in the progression of NSCLC, and suggested that targeting this pathway might be an effective therapeutic strategy in treating NSCLC.
我们最近的研究发现，QKI-5调控的微小RNA（miRNA），即miR-196b-5p，通过直接靶向GATA6、TSPAN12和FAS促进非小细胞肺癌（NSCLC）进展。然而，miR-196b-5p在NSCLC进展和转移中的生物学功能仍不清楚。
采用细胞增殖、迁移、集落形成、细胞周期分析来研究细胞表型变化。运用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹分析来检测相关基因和蛋白质的表达。通过RNA免疫沉淀（RIP）分析确定QKI-5与miR-196b-5p之间的相互作用。利用荧光素酶报告基因分析确定miR-196b-5p与NFKBIA 3'-非翻译区（3'-UTR）之间的直接结合。采用酶联免疫吸附测定（ELISA）检测分泌的白细胞介素6（IL6）蛋白。利用小鼠异种移植模型评估NFKBIA对体内肿瘤生长的作用。
我们证明，miR-196b-5p通过直接靶向NFKBIA（NFκB信号的负调节因子）促进肺癌细胞增殖、迁移、集落形成和细胞周期进程。敲低NFKBIA可增加IL6介导的信号转导和转录激活因子3（STAT3）磷酸化，从而通过激活NFκB信号促进肺癌细胞生长。NFKBIA的表达在NSCLC组织样本中显著下调，且与miR-196b-5p的表达呈负相关。此外，我们发现NSCLC中QKI-5表达下调与miR-224表达升高有关。
我们的研究结果表明，miR-224/QKI-5/miR-196b-5p/NFKBIA信号通路可能在NSCLC进展中发挥重要作用，并提示靶向该通路可能是治疗NSCLC的有效策略。
