Table 4_Inhibiting melanoma tumor growth: the role of oxidative stress-associated LINC02132 and COPDA1 long non-coding RNAs.xlsx

BackgroundCutaneous melanoma is a type of malignant tumor that is challenging to predict and is readily stimulated by various factors. Oxidative stress can induce damage and alterations in melanocytes, subsequently triggering immune responses. Given that oxidative stress is a prevalent tumor stimulus, we aimed to enhance melanoma prediction by identifying lncRNA signatures associated with oxidative stress.
MethodsWe screened for oxidative stress-related lncRNAs that could improve melanoma patient prognosis using the TCGA and GTEx databases. Utilizing differentially expressed oxidative stress-related lncRNAs (DE-OSlncRNAs), we constructed a Lasso regression model. The accuracy of the model was validated using univariate and multivariate regression, Kaplan-Meier (K-M) curves, and ROC curves. Subsequently, we conducted immune infiltration analysis, immune checkpoint differential analysis, IC50 pharmaceutical analysis, and gene set enrichment analysis. Investigating the effects of the target gene on melanoma using fluorescence in situ hybridization (FISH), quantitative real-time PCR (qRT-PCR), Edu assay, wound healing assay, transwell assay, flow cytometry, and reactive oxygen species (ROS) detection.
ResultsThirteen lncRNAs were identified as significant prognostic factors. Four oxidative stress-related lncRNAs (COPDA1, LINC02132, LINC02812, and MIR205HG) were further validated by fluorescence in situ hybridization (FISH), with results consistent with our data analysis. LINC02132 and COPDA1 can influence the proliferation, invasion, migration, and apoptosis of melanoma.
ConclusionOur findings suggest that upregulation of the LINC02132 or COPDA1 genes elevates intracellular reactive oxygen species (ROS) levels in melanoma cells, suppresses tumor cell proliferation, migration, and invasion, and promotes apoptosis. These results suggest a novel therapeutic strategy for melanoma treatment.

背景 皮肤黑色素瘤是一类预测难度较高且易受多种因素诱发的恶性肿瘤。氧化应激可诱导黑色素细胞发生损伤与表型改变，进而触发免疫应答。鉴于氧化应激是一类常见的肿瘤刺激因素，本研究旨在通过筛选与氧化应激相关的长链非编码RNA（lncRNA）特征，提升黑色素瘤的预测效能。
方法 本研究借助癌症基因组图谱（TCGA）与基因型组织表达数据库（GTEx），筛选可改善黑色素瘤患者预后的氧化应激相关lncRNA。通过差异表达氧化应激相关lncRNAs（DE-OSlncRNAs）构建Lasso回归模型，并采用单变量回归、多变量回归、卡普兰-迈耶（Kaplan-Meier，K-M）曲线以及受试者工作特征（ROC）曲线对模型的预测准确性进行验证。随后开展免疫浸润分析、免疫检查点差异分析、IC50药物敏感性分析以及基因集富集分析。本研究还通过荧光原位杂交（FISH）、实时定量聚合酶链反应（qRT-PCR）、EdU增殖实验、划痕愈合实验、Transwell实验、流式细胞术以及活性氧（ROS）检测，探究靶基因对黑色素瘤的调控作用。
结果 本研究共筛选出13个具有显著预后价值的lncRNA。其中4个氧化应激相关lncRNA（COPDA1、LINC02132、LINC02812及MIR205HG）通过荧光原位杂交（FISH）得到进一步验证，实验结果与数据分析结果一致。LINC02132与COPDA1可调控黑色素瘤细胞的增殖、侵袭、迁移及凋亡过程。
结论 本研究结果显示，上调LINC02132或COPDA1基因可提高黑色素瘤细胞内的活性氧（ROS）水平，抑制肿瘤细胞的增殖、迁移与侵袭，并促进细胞凋亡。上述研究结果为黑色素瘤的临床治疗提供了全新的潜在策略。