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| Effects of Bruceine D regulating the AMPK/ULK1 pathway on glycolysis in bladder cancer cells |
| Hits 29 Download times 2 Received:February 25, 2026 |
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| DOI
10.11656/j.issn.1672-1519.2026.05.10 |
| Key Words
Bruceine D;bladder cancer;proliferation;glycolysis;autophagy;apoptosis |
| Author Name | Affiliation | E-mail | | ZHANG Haibo | Department of Urology, People's Hospital of Zhengzhou, Zhengzhou 450003, China | | | GE Lei | Department of Urology, People's Hospital of Zhengzhou, Zhengzhou 450003, China | 875391450@qq.com | | CHEN Kun | Department of Urology, People's Hospital of Zhengzhou, Zhengzhou 450003, China | | | ZHANG Sen | Department of Urology, People's Hospital of Zhengzhou, Zhengzhou 450003, China | | | QIAN Xiaolei | Department of Urology, People's Hospital of Zhengzhou, Zhengzhou 450003, China | | | SHAN Zhongjie | Department of Urology, People's Hospital of Zhengzhou, Zhengzhou 450003, China | |
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| Abstract
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| [Objective] To investigate the effect of Bruceine D regulating the AMP-activated protein kinase（AMPK）/UNC-51-like autophagy-activated kinase 1（ULK1） pathway on glycolysis in bladder cancer cells. [Methods] The T24 cells were randomly divided into bladder cancer group，low-，medium-，and high-concentration Bruceine D groups，AMPK（AICAR activator） group，and high-concentration Bruceine D+AMPK inhibitor（Compound C） group. Cell proliferation was detected by clone formation assay and CCK-8 assay. Cell apoptosis was detected by flow cytometry. Kits were used to measure lactic acid production，glucose intake and amount of pyruvate production. The relative content of autophagosomes in cells was detected by monodansylcadaverine（MDC） staining. The mRNA levels of proliferating cell nuclear antigen（PCNA），Cleaved Caspase-3，pyruvate kinase M2（PKM2），and hexokinase 2（HK2） were measured by qRT-PCR. Western blot was used to examine the proteins of Beclin1，microtubule-associated protein 1 light chain 3（LC3）-Ⅱ/LC3-Ⅰ，p-AMPK/AMPK and p-ULK1/ULK1 in the cells. A nude mouse xenograft model of T24 cells was established to measure the effects of Bruceine D on the growth of xenograft tumors，and protein levels of AMPK/ULK1 pathway in tumor tissues by Western blot. [Results] Compared with the bladder cancer group，the number of T24 cell clone formation，OD450 value，lactic acid production，glucose uptake，amount of pyruvate production in the low-，medium-，and high-concentration Bruceine D groups and the AICAR grou were decreased，the mRNA levels of PCNA，PKM2，and HK2 were reduced，while the Cleaved Caspase-3 mRNA level was raised. The apoptosis rate，the relative content of autophagosomes，and the protein expressions of Beclin1，LC3-Ⅱ/LC3-Ⅰ，p-AMPK/AMPK，and p-ULK1/ULK1 were increased（P<0.05）. Compared with the high concentration Bruceine D group，the number of clone formation，OD450 value，lactic acid production，glucose uptake，amount of pyruvate production in the high concentration Bruceine D+Compound C group were added，PCNA，PKM2 and HK2 mRNA levels were increased，while Cleaved Caspase-3 mRNA level was reduced. The apoptosis rate，the relative content of autophagosomes，and the protein expressions of Beclin1，LC3-Ⅱ/LC3-Ⅰ，p-AMPK/AMPK，and p-ULK1/ULK1 were decreased（P<0.05）. Compared with the control group，the tumor tissue mass and volume in the Bruceine D group were decreased，while the p-AMPK/AMPK and p-ULK1/ULK1 protein levels in the tumor tissue were increased（P<0.05）. [Conclusion] Bruceine D may inhibit the proliferation of T24 cells，glycolysis，and the growth of xenograft tumors in nude mice，as well as induce cell autophagy and apoptosis，by activating the AMPK/ULK1 pathway. |
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