Abstract
目的
探究BCAT1在肺腺癌发展过程中的生物学作用及其分子机制。
创新点
首次证实BCAT1通过调控线粒体功能和NF-κB通路影响肺腺癌发展,为肺腺癌发展的分子机制研究和临床治疗提供了新见解。
方法
首先通过免疫组化确定BCAT1蛋白在临床肺腺癌患者癌与癌旁组织中的表达,然后采用慢病毒侵染法构建BCAT1过表达或敲低的A549稳转株并进行增殖、迁移和侵袭实验。此外分别通过裸鼠皮下成瘤实验研究BCAT1对肺腺癌体内成瘤与肿瘤生长能力的影响;通过CCK8、qRT-PCR、活性氧与氧消耗率检测等实验研究BCAT1对支链氨基酸代谢和线粒体功能的影响。最后经转录组学与蛋白质免疫印迹(Western blot)筛选并确定BCAT1影响肺腺癌发展的信号通路。
结论
BCAT1蛋白在肺腺癌患者的肺癌组织中呈过高表达状态,且BCAT1可增强肺腺癌细胞在体外与体内的增殖与迁移侵袭能力。BCAT1蛋白可能通过增强肺腺癌细胞中支链氨基酸代谢、线粒体生物合成和呼吸作用,降低胞内ROS含量,下调NFKBIB转录水平并激活NF-κB通路,促进肺腺癌发展。
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Availability of data
The RNA sequencing data were submitted to BioSample database of NCBI (BioProject ID: PRJNA717911).
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Acknowledgments
This work was supported by the National Science and Technology Major Project of China (No. 2018ZX10302205), the Major Project of Science and Technology in Henan Province (No. 161100311400), the Natural Science Foundation of Henan Province (Nos. 182300410361 and 222300420306), the Project of Basic Research Fund of Henan Institute of Medical and Pharmacological Sciences (No. 2022BP0103), and the Key Scientific and Technological Project of Henan Province (Nos. 212102310124 and 222102310083). We appreciate the continuous support of Henan Key Laboratory for Pharmacology of Liver Diseases where most of the experiments were performed.
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Qianwei ZHAO, Jintao ZHANG, and Jianying ZHANG designed the study. Mengdan YU, Jinxia LI, and Zhibiao ZHANG performed the experiments and analyzed data. Fang XU, Yixian LIU, Liping DAI, and Bingxia ZHANG collected materials and helped in manuscript preparing. Qianwei ZHAO, Mengdan YU, and Jinxia LI wrote the manuscript. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Detailed methods are provided in the electronic supplementary materials of this paper.
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Mengdan YU, Qianwei ZHAO, Jinxia LI, Fang XU, Zhibiao ZHANG, Yixian LIU, Liping DAI, Bingxia ZHANG, Jianying ZHANG, and Jintao ZHANG declare that they have no conflict of interest.
All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008 (5). Informed consent was obtained from all patients for being included in the study. All institutional and national guidelines for the care and use of laboratory animals were followed.
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Material and methods; Figs. S1–S5
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Yu, M., Zhao, Q., Li, J. et al. BCAT1 promotes lung adenocarcinoma progression through enhanced mitochondrial function and NF-κB pathway activation. J. Zhejiang Univ. Sci. B 23, 760–769 (2022). https://doi.org/10.1631/jzus.B2100985
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DOI: https://doi.org/10.1631/jzus.B2100985