乳酸通过调节 m6A 修饰介导的 FOXO3/MAGI1-IT1/miR-664b-3p/IL-6R 轴促进 NSCLC 顺铂耐药。
Lactate promoted cisplatin resistance in NSCLC by modulating the m6A modification-mediated FOXO3/MAGI1-IT1/miR-664b-3p/IL-6R axis.
机构信息
College of Integrated Chinese and Western Medical, Liaoning University of Traditional Chinese Medicine, No. 79 Chongshan, Huanggu District, Shenyang, Liaoning 118047, China; Pathology Department of Shenyang Medical College, Shenyang, Liaoning 110034, China.
College of Integrated Chinese and Western Medical, Liaoning University of Traditional Chinese Medicine, No. 79 Chongshan, Huanggu District, Shenyang, Liaoning 118047, China.
出版信息
Neoplasia. 2024 Feb;48:100960. doi: 10.1016/j.neo.2023.100960. Epub 2024 Jan 6.
BACKGROUND
Cisplatin resistance is one of the major obstacles in non-small cell lung cancer (NSCLC) treatment. Intriguingly, elevated lactate levels were observed in cisplatin-resistant cells, which spurred further investigation into their underlying biological mechanisms.
METHODS
Lactate levels were measured by lactate detection kit. Cisplatin-resistance NSCLC cells were established using progressive concentration of cisplatin. Cell viability, proliferation, and apoptosis were detected by CCK-8, EdU, and flow cytometry, respectively. Cell proliferation in vivo was determined by immunohistochemistry of Ki67 and apoptotic cells were calculated by the TUNEL. MeRIP-PCR was used to measure FOXO3 m6A levels. The interactions of genes were analyzed via RIP, ChIP, Dual-luciferase reporter, and RNA pull-down, respectively.
RESULTS
Elevated lactate levels were observed in both NSCLC patients and cisplatin-resistance cells. Lactate treatment increased cisplatin-resistance cell viability in vitro and promoted tumor growth in vivo. Mechanistically, lactate downregulated FOXO3 by YTHDF2-mediated m6A modification. FOXO3 transcriptionally reduced MAGI1-IT1 expression. FOXO3 overexpression inhibited the lactate-induced promotion of cisplatin resistance in NSCLC, which were reversed by MAGI1-IT1 overexpression. MAGI1-IT1 and IL6R competitively bound miR-664b-3p. FOXO3 overexpression or MAGI1-IT1 knockdown repressed lactate-mediated cisplatin resistance in vivo.
CONCLUSION
Lactate promoted NSCLC cisplatin resistance through regulating FOXO3/MAGI1-IT1/miR-664b-3p/IL6R axis in YTHDF2-mediated m6A modification.
背景
顺铂耐药是非小细胞肺癌（NSCLC）治疗的主要障碍之一。有趣的是，在顺铂耐药细胞中观察到升高的乳酸水平，这促使进一步研究其潜在的生物学机制。
方法
通过乳酸检测试剂盒测量乳酸水平。使用顺铂的逐渐浓度建立顺铂耐药 NSCLC 细胞。通过 CCK-8、EdU 和流式细胞术分别检测细胞活力、增殖和凋亡。通过 Ki67 的免疫组化测定体内细胞增殖，通过 TUNEL 计算凋亡细胞。使用 MeRIP-PCR 测量 FOXO3 的 m6A 水平。通过 RIP、ChIP、双荧光素酶报告基因和 RNA 下拉分别分析基因的相互作用。
结果
在 NSCLC 患者和顺铂耐药细胞中均观察到升高的乳酸水平。乳酸处理增加了体外顺铂耐药细胞的活力，并促进了体内肿瘤的生长。在机制上，乳酸通过 YTHDF2 介导的 m6A 修饰下调 FOXO3。FOXO3 转录下调 MAGI1-IT1 的表达。FOXO3 过表达抑制 NSCLC 中乳酸诱导的顺铂耐药，而 MAGI1-IT1 过表达则逆转了这一作用。MAGI1-IT1 和 IL6R 竞争性结合 miR-664b-3p。FOXO3 过表达或 MAGI1-IT1 敲低抑制体内乳酸介导的顺铂耐药。
结论
乳酸通过 YTHDF2 介导的 m6A 修饰调节 FOXO3/MAGI1-IT1/miR-664b-3p/IL6R 轴促进 NSCLC 顺铂耐药。
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