近期研究报告显示，昼夜节律转录因子芳香族碳氢化合物受体核转移因子2（ARNTL2）促进肺腺癌的转移进展。然而，ARNTL2在非小细胞肺癌（NSCLC）细胞生长和增殖中的分子机制尚待探索。在TCGA数据库中分析肺癌患者的ARNTL2和酰基辅酶A硫酯酯酶7（ACOT7）表达。通过转染细胞质粒或慢病毒来进行ARNTL2和ACOT7的功能增强。通过siRNA进行Knockdown实验。使用Western blot和qRT-PCR检查蛋白质和mRNA表达情况。使用Dual luciferase和ChIP-qPCR检测ARNTL2对ACOT7的启动子序列的相互作用。使用指示性检测工具来测量三酰甘油水平、MDA产生量、Caspase 3到Caspase 7的活性和脂质ROS。使用CCK8、集落形成和流式细胞术分析细胞死亡和细胞周期检测细胞功能。我们证明了ARNTL2上调ACOT7对NSCLC细胞生长和增殖至关重要。首先，ARNTL2过表达与LUAD患者的不良预后相关，并支持NSCLC细胞的增殖。通过分子实验，我们表明，ARNTL2通过直接结合到ACOT7的启动子序列强化了ACOT7基因的转录活性。ACOT7高表达与LUAD患者的预后越差相关。功能增强和功能缺陷实验揭示了AOCT7对NSCLC细胞生长和增殖的贡献。ACOT7调节了NSCLC细胞的凋亡和铁死亡，但对细胞周期进展没有影响。ACOT7过表达也增强了脂肪酸合成并抑制了脂质过氧化反应。最后，我们展示ARNTL2敲减和过表达抑制和促进细胞三酰甘油的生产和随后的细胞增殖，这可以被ACOT7过表达和敲降所逆转。我们的研究阐明了ARNTL2 / ACOT7轴在NSCLC发展中的致癌功能。针对ARNTL2或ACOT7可能是具有高表达ARNTL2的NSCLC患者的有前途的治疗策略。2023年，作者（们）。

Recent studies have reported that the circadian transcription factor aryl hydrocarbon receptor nuclear translocator like 2 (ARNTL2) promotes the metastatic progression of lung adenocarcinoma. However, the molecular mechanisms of ARNTL2 in non-small cell lung cancer (NSCLC) cell growth and proliferation remain to be explored.The expression of ARNTL2 and acyl-CoA thioesterase 7 (ACOT7) in lung cancer patients was analyzed based on TCGA database. Gain-of-function of ARNTL2 and ACOT7 was conducted by transfecting the cells with plasmids or lentivirus. Knockdown assay was carried out by siRNAs. Western blot and qRT-PCR were performed to check the protein and mRNA expression. Dual luciferase and ChIP-qPCR assay was applied to check the interaction of ARNTL2 on ACOT7's promoter sequence. Triglyceride level, MDA production, the activity of casapase 3 to caspase 7, and lipid ROS were measured by indicated assay kit. Cellular function was detected by CCK8, colony formation and flow cytometry analysis of cell death and cell cycle.We demonstrated that ARNTL2 upregulation of ACOT7 was critical for NSCLC cell growth and proliferation. Firstly, overexpression of ARNTL2 conferred the poor prognosis of LUAD patients and supported the proliferation of NSCLC cells. Based on molecular experiments, we showed that ARNTL2 potentiated the transcription activity of ACOT7 gene via direct binding to ACOT7's promoter sequence. ACOT7 high expression was correlated with the worse prognosis of LUAD patients. Gain-of-function and loss-of-function experiments revealed that AOCT7 contributed to NSCLC cell growth and proliferation. ACOT7 regulated the apoptosis and ferroptosis of NSCLC cells, while exhibited no effect on cell cycle progression. ACOT7 overexpression also potentiated fatty acid synthesis and suppressed lipid peroxidation. Lastly, we showed that ARNTL2 knockdown and overexpression inhibited and promoted the cellular triglyceride production and subsequent cell proliferation, which could be reversed by ACOT7 overexpression and knockdown.Our study illustrated the oncogenic function of ARNTL2/ACOT7 axis in the development of NSCLC. Targeting ARNTL2 or ACOT7 might be promising therapeutic strategies for NSCLC patients with highly expressed ARNTL2.© 2023. The Author(s).