Ubiquitin-conjugating enzyme E2T (UBE2T) acts as an oncogene in various types of cancer. However, the mechanisms behind its oncogenic role remain unclear in lung cancer. This study aims to explore the function and clinical relevance of UBE2T in lung cancer. Lentiviral vectors were used to mediate UBE2T depletion or overexpress UBE2T in lung cancer cells. CCK8 analysis and western blotting were performed to investigate the effects of UBE2T on proliferation, autophagy, and relevant signaling pathways. To exploit the clinical significance of UBE2T, we performed immunohistochemistry staining with an anti-UBE2T antibody on 131 NSCLC samples. Moreover, we downloaded the human lung adenocarcinoma (LUAD) dataset from The Cancer Atlas Project (TCGA). Lasso Cox regression model was adopted to establish a prognostic model with UBE2T-correlated autophagy genes. We found that UBE2T stimulated proliferation and autophagy, and silencing this gene abolished autophagy in lung cancer cells. As suggested by Gene set enrichment analysis, we observed that UBE2T downregulated p53 levels in A549 cells and vice versa. Blockade of p53 counteracted the inhibitory effects of UBE2T depletion on autophagy. Meanwhile, the AMPK/mTOR signaling pathway was activated during UBE2T-mediated autophagy, suggesting that UBE2T promotes autophagy via the p53/AMPK/mTOR pathway. Interestingly, UBE2T overexpression increased cisplatin-trigged autophagy and led to cisplatin resistance of A549 cells, whereas inhibiting autophagy reversed drug resistance. However, no association was observed between UEB2T and overall survival in a population of 131 resectable NSCLC patients. Therefore, we developed and validated a multiple gene signature by considering UBE2T and its relevance in autophagy in lung cancer. The risk score derived from the prognostic signature significantly stratified LUAD patients into low- and high-risk groups with different overall survival. The risk score might independently predict prognosis. Interestingly, nomogram and decision curve analysis demonstrated that the signature’s prognostic accuracy culminated while combined with clinical features. Finally, the risk score showed great potential in predicting clinical chemosensitivity. We found that UBE2T upregulates autophagy in NSCLC cells by activating the p53/AMPK/mTOR signaling pathway. The clinical predicting ability of UBE2T in LUAD can be improved by considering the autophagy-regulatory role of UBE2T.

中文翻译：

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UBE2T通过p53/AMPK/mTOR信号通路促进肺腺癌自噬

泛素结合酶 E2T (UBE2T) 在各种类型的癌症中充当癌基因。然而，其致癌作用背后的机制在肺癌中仍不清楚。本研究旨在探讨UBE2T在肺癌中的功能和临床相关性。慢病毒载体用于介导肺癌细胞中的 UBE2T 消耗或过度表达 UBE2T。进行 CCK8 分析和蛋白质印迹以研究 UBE2T 对增殖、自噬和相关信号通路的影响。为了利用 UBE2T 的临床意义，我们用抗 UBE2T 抗体对 131 个 NSCLC 样本进行了免疫组织化学染色。此外，我们从癌症图谱项目 (TCGA) 下载了人类肺腺癌 (LUAD) 数据集。采用 Lasso Cox 回归模型建立 UBE2T 相关自噬基因的预后模型。我们发现 UBE2T 刺激增殖和自噬，沉默该基因消除了肺癌细胞中的自噬。正如基因集富集分析所建议的，我们观察到 UBE2T 下调 A549 细胞中的 p53 水平，反之亦然。p53 的阻断抵消了 UBE2T 消耗对自噬的抑制作用。同时，AMPK/mTOR 信号通路在 UBE2T 介导的自噬过程中被激活，表明 UBE2T 通过 p53/AMPK/mTOR 通路促进自噬。有趣的是，UBE2T 过表达增加了顺铂引发的自噬并导致 A549 细胞对顺铂产生耐药性，而抑制自噬则逆转了耐药性。然而，在 131 名可切除 NSCLC 患者中，未观察到 UEB2T 与总生存期之间存在关联。因此，我们通过考虑 UBE2T 及其与肺癌自噬的相关性开发并验证了多基因特征。来自预后特征的风险评分显着将 LUAD 患者分层为具有不同总生存期的低风险和高风险组。风险评分可能独立预测预后。有趣的是，列线图和决策曲线分析表明，特征的预后准确性在与临床特征相结合时达到顶峰。最后，风险评分在预测临床化学敏感性方面显示出巨大的潜力。我们发现 UBE2T 通过激活 p53/AMPK/mTOR 信号通路上调 NSCLC 细胞中的自噬。