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目的 探究基质硬度调控肝癌肿瘤再生细胞（TRCs）干性及恶性表型的分子机制，明确果糖二磷酸醛缩酶C（ALDOC）在力学微环境介导的糖代谢重编程中的作用。方法 构建90、420、1 050Pa3种硬度的三维纤维蛋白凝胶模拟体内力学微环境，筛选适宜肝癌TRCs生长的软基质条件；采用克隆形成实验检测不同硬度基质对HepG2细胞克隆体积的影响，通过Transwell实验评估细胞侵袭转移能力，利用RT-qPCR检测干性基因及糖酵解相关酶的表达水平；敲低或过表达ALDOC，结合葡萄糖、乳酸检测试剂盒分析细胞糖酵解水平变化。结果 90Pa软基质组肝癌TRCs克隆体积显著大于420、1 050Pa硬基质组，且差异随培养时间延长而增大（P<0.001）；软基质诱导的TRCs侵袭转移能力增强（P<0.01），干性基因NANOG、OCT4等表达水平显著上调；转录组测序及RT-qPCR验证显示，软基质可广泛上调糖酵解相关酶表达，其中ALDOC上调最为显著；敲低ALDOC后，肝癌TRCs葡萄糖摄取量降低、乳酸生成量减少，克隆增殖能力显著下降，而过表达ALDOC则逆转上述表型。结论 软基质可通过上调ALDOC表达介导糖酵解代谢重编程，进而维持肝癌TRCs干性。
Abstract：Objective To investigate the molecular mechanisms by which matrix stiffness regulates the stemness and malignant phenotypes of hepatocellular carcinoma tumor repopulating cells (TRCs)， elucidate the role of aldolase C (ALDOC) in mechano-microenvironment-mediated reprogramming of glucose metabolism. Methods Three-dimensional fibrin gels with stiffnesses of 90 ， 420 ， and 1 050 Pa were constructed to simulate the in vivo mechanical microenvironment， and identify the optimal substrate stiffness for hepatocellular carcinoma TRCs growth. Colony formation assays assessed the effects of different matrix stiffnesses on HepG2 colony volumes. Transwell assays evaluated cell invasion and migration capabilities. RT-qPCR assays measured mRNA expression levels of stemness genes and glycolysis-related enzymes. ALDOC expression was knocked down via small interfering RNA transfection， and cellular glycolysis levels were analyzed using glucose and lactate detection kits. Results Hepatocellular carcinoma TRCs cultured in the 90 Pa soft substrate formed significantly larger colonies volumes compared to those in the 420 Pa and 1 050 Pa stiff substrates， and the difference increased with extended culture time (P<0.001). The soft substrate markedly enhanced the invasive and migratory capacities of TRCs (P<0.01) and upregulated the expression of stemness-related genes such as NANOG and OCT4. Transcriptome sequencing and RT-qPCR assays revealed that soft matrix broadly upregulated the expression of glycolysis-related enzymes， with ALDOC showing the most significant increase. Following ALDOC knockdown， hepatocellular carcinoma TRCs exhibited reduced glucose uptake and lactate production， accompanied by a marked decline in proliferation capacity. In contrast， overexpression of ALDOC reversed the aforementioned phenotype. Conclusion Soft matrix mediates glycolytic metabolic reprogramming by upregulating ALDOC expression， thereby maintaining hepatocellular carcinoma TRCs stemness.
[1] Sung H， Ferlay J， Siegel R L， et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin， 2021， 71(3): 209-49. doi:10.3322/caac.21660.
[2] Hong J， Yu J， Buratto D， et al. Unveiling the role of mechanical microenvironment in hepatocellular carcinoma: molecular mechanisms and implications for therapeutic strategies[J]. Int J Biol Sci， 2024， 20(13): 5239-53. doi:10.7150/ijbs.102706.
[3] Fu X， Zhang Y， Luo Q， et al. Targeting the mechano-microenvironment and liver cancer stem cells: a promising therapeutic strategy for liver cancer[J]. Cancer Biol Med， 2023: 1-14. doi:10.20892/j.issn.2095-3941.2023.0229.
[4] Sistigu A， Musella M， Galassi C， et al. Tuning cancer fate: tumor microenvironment’s role in cancer stem cell quiescence and reawakening[J]. Front Immunol， 2020， 11: 2166. doi:10.3389/fimmu.2020.02166.
[5] Tan Y， Tajik A， Chen J， et al. Matrix softness regulates plasticity of tumour-repopulating cells via H3K9 demethylation and Sox2 expression[J]. Nat Commun， 2014， 5: 4619. doi:10.1038/ncomms5619.
[6] Yadav U P， Singh T， Kumar P， et al. Metabolic adaptations in cancer stem cells[J]. Front Oncol， 2020， 10: 1010. doi:10.3389/fonc.2020.01010.
[7] 张 黎， 崔 杰， 王安琪， 等. ATF4通过调控HKDC1对肝癌细胞增殖、迁移及凋亡的影响[J]. 安徽医科大学学报， 2021， 56(12): 1932-8. doi:10.19405/j.cnki.issn1000-1492.2021.12.016.
[7] Zhang L， Cui J， Wang A Q， et al. Effect of ATF4 on the proliferation， migration and apoptosis of hepatocellular carcinoma by regulating HKDC1 protein[J]. Acta Univ Med Anhui， 2021， 56(12): 1932-8. doi:10.19405/j.cnki.issn1000-1492.2021.12.016.
[8] Park J S， Burckhardt C J， Lazcano R， et al. Mechanical regulation of glycolysis via cytoskeleton architecture[J]. Nature， 2020， 578(7796): 621-6. doi:10.1038/s41586-020-1998-1.
[9] Sohrabi A， Lefebvre A E Y T， Harrison M J， et al. Microenvironmental stiffness induces metabolic reprogramming in glioblastoma[J]. Cell Rep， 2023， 42(10): 113175. doi:10.1016/j.celrep.2023.113175.
[10] Zheng J H， Zhu Y H， Yang J， et al. A CLIC1 network coordinates matrix stiffness and the Warburg effect to promote tumor growth in pancreatic cancer[J]. Cell Rep， 2024， 43(8): 114633. doi:10.1016/j.celrep.2024.114633.
[11] DuFort C C， Paszek M J， Weaver V M. Balancing forces: architectural control of mechanotransduction[J]. Nat Rev Mol Cell Biol， 2011， 12(5): 308-19. doi:10.1038/nrm3112.
[12] Tang K， Zheng Y， Hu G， et al. Local soft niches in mechanically heterogeneous primary tumors promote brain metastasis via mechanotransduction-mediated HDAC3 activity[J]. Sci Adv， 2025， 11(9): eadq2881. doi:10.1126/sciadv.adq2881.
[13] Niu Y， Lin Z， Wan A， et al. Loss-of-function genetic screening identifies aldolase a as an essential driver for liver cancer cell growth under hypoxia[J]. Hepatology， 2021， 74(3): 1461-79. doi:10.1002/hep.31846.
[14] Li M， He X， Guo W， et al. Aldolase B suppresses hepatocellular carcinogenesis by inhibiting G6PD and pentose phosphate pathways[J]. Nat Cancer， 2020， 1(7): 735-47. doi:10.1038/s43018-020-0086-7.
基本信息:
中图分类号:R735.7
引用信息:
[1]项阳,张满意,张姚.软基质通过上调ALDOC增强糖酵解维持肝癌干性[J].安徽医科大学学报().
基金信息:
国家自然科学基金项目（编号：12502359）; 安徽医科大学基础与临床合作研究提升计划项目（编号：2023xkjT015）
2026-08-31
2026-08-31
2026-08-31