Sun Ke-Xiang, Tan Wei-Shan, Wang Hao-Yue, Gao Jia-Min, Wang Shu-Yun, Xie Man-Li, Deng Wan-Li
Department of Medical Oncology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200062, China.
Chin J Integr Med. 2025 Jun;31(6):529-540. doi: 10.1007/s11655-024-4113-x. Epub 2024 Nov 25.
To explore the role of the natural compound hesperidin in glycolysis, the key ratelimiting enzyme, in colorectal cancer (CRC) cell lines.
In vitro, HCT116 and SW620 were treated with different doses of hesperidin (0-500 µmol/L), cell counting kit-8 and colone formation assays were utilized to detected inhibition effect of hesperidin on CRC cell lines. Transwell and wound healing assays were performed to detect the ability of hesperidin (0, 25, 50 and 75 µmol/L) to migrate CRC cells. To confirm the apoptotic-inducing effect of hesperidin, apoptosis and cycle assays were employed. Western blot, glucose uptake, and lactate production determination measurements were applied to determine inhibitory effects of hesperidin (0, 25 and 50 µmol/L) on glycolysis. In vivo, according to the random number table method, nude mice with successful tumor loading were randomly divided into vehicle, low-dose hesperidin (20 mg/kg) and high-dose hesperidin (60 mg/kg) groups, with 6 mice in each group. The body weights and tumor volumes of mice were recorded during 4-week treatment. The expression of key glycolysis rate-limiting enzymes was determined using Western blot, and glucose uptake and lactate production were assessed. Finally, protein interactions were probed with DirectDIA Quantitative Proteomics, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses.
Hesperidin could inhibit CRC cell line growth (P<0.05 or P<0.01). Moreover, hesperidin presented an inhibitory effect on the migrating abilities of CRC cells. Hesperidin also promoted apoptosis and cell cycle alterations (P<0.05). The immunoblotting results manifested that hesperidin decreased the levels of hexokinase 2, glucose transporter protein 1 (GLUT1), GLUT3, L-lactate dehydrogenase A, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2 (PFKFB2), PFKFB3, and pyruvate kinase isozymes M2 (P<0.01). It remarkably suppressed tumor xenograft growth in nude mice. GO and KEGG analyses showed that hesperidin treatment altered metabolic function.
Hesperidin inhibits glycolysis and is a potential therapeutic choice for CRC treatment.
探讨天然化合物橙皮苷对结直肠癌（CRC）细胞系中糖酵解关键限速酶的作用。
在体外，用不同剂量的橙皮苷（0 - 500 μmol/L）处理HCT116和SW620细胞，采用细胞计数试剂盒-8和克隆形成试验检测橙皮苷对CRC细胞系的抑制作用。进行Transwell和伤口愈合试验，检测橙皮苷（0、25、50和75 μmol/L）对CRC细胞迁移能力的影响。为证实橙皮苷的诱导凋亡作用，采用凋亡和细胞周期试验。应用蛋白质免疫印迹法、葡萄糖摄取和乳酸生成测定，以确定橙皮苷（0、25和50 μmol/L）对糖酵解的抑制作用。在体内，按照随机数字表法，将成功荷瘤的裸鼠随机分为溶剂对照组、低剂量橙皮苷（20 mg/kg）组和高剂量橙皮苷（60 mg/kg）组，每组6只小鼠。在4周的治疗过程中记录小鼠的体重和肿瘤体积。采用蛋白质免疫印迹法测定关键糖酵解限速酶的表达，并评估葡萄糖摄取和乳酸生成情况。最后，通过DirectDIA定量蛋白质组学、基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析探索蛋白质相互作用。
橙皮苷可抑制CRC细胞系生长（P<0.05或P<0.01）。此外，橙皮苷对CRC细胞的迁移能力有抑制作用。橙皮苷还可促进细胞凋亡和细胞周期改变（P<0.05）。免疫印迹结果表明，橙皮苷可降低己糖激酶2、葡萄糖转运蛋白1（GLUT1）、GLUT3、L-乳酸脱氢酶A、6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶2（PFKFB2）、PFKFB3和丙酮酸激酶同工酶M2的水平（P<0.01）。它能显著抑制裸鼠体内肿瘤异种移植的生长。GO和KEGG分析表明，橙皮苷处理可改变代谢功能。
橙皮苷抑制糖酵解，是CRC治疗的潜在选择。
