Song Gaojie, Fang Jinbo, Shang Chao, Li Yiquan, Zhu Yilong, Xiu Zhiru, Sun Lili, Jin Ningyi, Li Xiao
Medical College, Yanbian University, Yanji, China; Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, China; Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun, China.
Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun, China.
Exp Cell Res. 2021 Dec 15;409(2):112926. doi: 10.1016/j.yexcr.2021.112926. Epub 2021 Nov 15.
Ad-apoptin is a recombinant oncolytic adenovirus constructed by our laboratory that can express apoptin. It can selectively kill tumor cells without damaging normal cells. This study investigated the effects of Ad-apoptin on glycolysis, migration and invasion of non-small cell lung cancer. Cell viability and apoptosis were detected by CCK-8 and flow cytometry, respectively. Glycolysis was investigated by glucose consumption, lactic acid production and glycolytic key enzyme protein levels. Migration and invasion were evaluated via wound healing, transwell assays and epithelial-mesenchymal transition (EMT) protein levels. The interaction between apoptin and AMPK was detected by Co-IP. A nude mice tumor model was established to investigate the anti-cancer role of Ad-apoptin in vivo. The results showed that Ad-apoptin inhibits cell viability and induces apoptosis of A549 and NCI-H23 cells. Ad-apoptin can reduce the glucose uptake and lactic production in lung cancer cells, and reduce the expression of related glycolysis-limiting enzymes. At the same time, Ad-apoptin inhibited the migration and invasion of lung cancer. Immunoprecipitation showed that apoptin and AMPK could interact directly. Moreover, knockdown of AMPK significantly attenuated the inhibitory effect of Ad-apoptin on glycolysis, migration and invasion of A549 and NCI-H23 cells. Ad-apoptin can inhibit the growth of tumors in nude mice. Compared with the control group, Ad-apoptin had a significant inhibitory effect on AMPK knockdown tumors. The immunohistochemical results of tumor tissues were consistent with those in vitro. Collectively, Ad-apoptin targets AMPK and inhibits glycolysis, migration and invasion of lung cancer cells through the AMPK/mTOR signaling pathway. This suggests that Ad-apoptin may have therapeutic potential for lung cancer by targeting AMPK activation.
Ad-apoptin是我们实验室构建的一种能表达凋亡素的重组溶瘤腺病毒。它能选择性地杀死肿瘤细胞而不损伤正常细胞。本研究探讨了Ad-apoptin对非小细胞肺癌糖酵解、迁移和侵袭的影响。分别采用CCK-8法和流式细胞术检测细胞活力和凋亡情况。通过葡萄糖消耗、乳酸生成和糖酵解关键酶蛋白水平来研究糖酵解。通过伤口愈合实验、Transwell实验和上皮-间质转化（EMT）蛋白水平评估迁移和侵袭能力。通过免疫共沉淀检测凋亡素与AMPK之间的相互作用。建立裸鼠肿瘤模型以研究Ad-apoptin在体内的抗癌作用。结果表明，Ad-apoptin抑制A549和NCI-H23细胞的活力并诱导其凋亡。Ad-apoptin可降低肺癌细胞的葡萄糖摄取和乳酸生成，并降低相关糖酵解限速酶的表达。同时，Ad-apoptin抑制肺癌的迁移和侵袭。免疫沉淀显示凋亡素与AMPK可直接相互作用。此外，敲低AMPK可显著减弱Ad-apoptin对A549和NCI-H23细胞糖酵解、迁移和侵袭的抑制作用。Ad-apoptin可抑制裸鼠体内肿瘤的生长。与对照组相比，Ad-apoptin对敲低AMPK的肿瘤具有显著抑制作用。肿瘤组织的免疫组化结果与体外实验结果一致。总体而言，Ad-apoptin靶向AMPK并通过AMPK/mTOR信号通路抑制肺癌细胞的糖酵解、迁移和侵袭。这表明Ad-apoptin通过靶向激活AMPK可能对肺癌具有治疗潜力。
