Li Yanguang, Niu Jieting, Sun Zhiguang, Liu Junfeng
Department of thoracic surgery, Cangzhou Central Hospital, Cangzhou City, Hebei Province, China.
Department of geriatric internal medicine, Cangzhou Central Hospital, Cangzhou City, Hebei Province, China.
J Biochem Mol Toxicol. 2025 Apr;39(4):e70232. doi: 10.1002/jbt.70232.
Potassium voltage-gated channel subfamily A regulatory beta subunit 2 (KCNAB2) is a potassium voltage-gated channel subfamily A member that plays a role in non-small cell lung cancer (NSCLC). However, its functional impact and mechanism in NSCLC are not fully understood. Here, we analyzed its effects on NSCLC cell behaviors and the underlying mechanism.mRNA expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR),(qRT-PCR), while protein expression was quantified by western blotting blot analysis or immunohistochemistry assay. NSCLC cell proliferation, migration, invasion, macrophage polarization, and apoptosis were evaluated through cell-based assays including cell counting kit-8 (CCK-8)(CCK-8) assay, flow cytometry, Tunel assay, wound-healing assay, and transwell invasion assay. The role of FTO alpha-ketoglutarate dependent dioxygenase (FTO)-mediated(FTO)-mediated m6A methylation in the regulation of KCNAB2 expression and their impacts on NSCLC cell behavior and M2 macrophage polarization were assessed through m6A RNA immunoprecipitation assay and rescue experiments. Xenograft mouse model assay was used to determine the effect of KCNAB2 on tumor formation in vivo.in vivo.KCNAB2 expression was downregulated and FTO expression was upregulated in NSCLC tissues and cells when compared with controls. Moreover, the expression of KCNAB2 was found to be lower in stage III NSCLC patients compared to those at stages I and II, and it was also lower in patients with positive lymph node metastasis compared to those with negative lymph node metastasis. Overexpression of KCNAB2 inhibited NSCLC cell proliferation, migration, invasion, and M2 macrophage polarization, while inducing cell apoptosis. These effects were mediated, at least partially, by inactivating the phosphoinositide 3-kinase (PI3K)/AKT(PI3K)/AKT pathway. Moreover, ectopic expression of KCNAB2 delayed tumor formation in vivo. FTOin vivo. FTO was found to mediate m6A methylation of KCNAB2, and knockdown of FTO resulted in the upregulation of KCNAB2 expression, leading to inhibition of NSCLC cell behavior and M2 macrophage polarization.KCNAB2 overexpression inhibited NSCLC cell behavior and M2 macrophage polarization by inactivating the PI3KPI3K/AKT/AKT pathway. Furthermore, FTOFTO-mediated-mediated m6A methylation was involved in the regulation of KCNAB2 expression in NSCLC. These results enhance our understanding of the role of KCNAB2 in NSCLC and suggest its potential as a therapeutic target.
钾电压门控通道亚家族A调节β亚基2（KCNAB2）是钾电压门控通道亚家族A的成员，在非小细胞肺癌（NSCLC）中发挥作用。然而，其在NSCLC中的功能影响和机制尚未完全阐明。在此，我们分析了其对NSCLC细胞行为的影响及潜在机制。通过定量实时聚合酶链反应（qRT-PCR）检测mRNA表达水平，而蛋白质表达则通过蛋白质免疫印迹分析或免疫组织化学测定进行定量。通过基于细胞的检测方法，包括细胞计数试剂盒-8（CCK-8）检测、流式细胞术、Tunel检测、伤口愈合检测和Transwell侵袭检测，评估NSCLC细胞的增殖、迁移、侵袭、巨噬细胞极化和凋亡。通过m6A RNA免疫沉淀检测和挽救实验，评估FTOα-酮戊二酸依赖性双加氧酶（FTO）介导的m6A甲基化在KCNAB2表达调控中的作用及其对NSCLC细胞行为和M2巨噬细胞极化的影响。采用异种移植小鼠模型实验来确定KCNAB2对体内肿瘤形成的影响。与对照相比，NSCLC组织和细胞中KCNAB2表达下调，FTO表达上调。此外，发现III期NSCLC患者中KCNAB2的表达低于I期和II期患者，并且淋巴结转移阳性患者中KCNAB2的表达也低于淋巴结转移阴性患者。KCNAB2的过表达抑制了NSCLC细胞的增殖、迁移、侵袭和M2巨噬细胞极化，同时诱导细胞凋亡。这些作用至少部分是通过使磷酸肌醇3-激酶（PI3K）/AKT信号通路失活介导的。此外，KCNAB2的异位表达在体内延迟了肿瘤形成。发现FTO介导KCNAB2的m6A甲基化，敲低FTO导致KCNAB2表达上调，从而抑制NSCLC细胞行为和M2巨噬细胞极化。KCNAB2过表达通过使PI3K/AKT信号通路失活来抑制NSCLC细胞行为和M2巨噬细胞极化。此外，FTO介导的m6A甲基化参与了NSCLC中KCNAB2表达的调控。这些结果加深了我们对KCNAB2在NSCLC中作用的理解，并表明其作为治疗靶点的潜力。
