哇巴因通过G6PD通路调节糖酵解影响结肠癌细胞增殖的机制研究

徐建雄; 努尔比亚木·达吾提; 萨尼耶·依不拉音; 帕孜来提; 李欣哲; 李文芳

doi:10.13568/j.cnki.651094.651316.2024.08.21.0001

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哇巴因通过G6PD通路调节糖酵解影响结肠癌细胞增殖的机制研究

结肠癌药物研究专题 | 更新时间：2026-01-21

- 哇巴因通过G6PD通路调节糖酵解影响结肠癌细胞增殖的机制研究
- 哇巴因通过G6PD通路调节糖酵解影响结肠癌细胞增殖的机制研究
- 新疆大学学报（自然科学版中英文） 2025年42卷第5期页码：621-631
- 作者机构：
  
  1. 新疆大学药学院(药物研究所)
  2. 新疆大学生命科学与技术学院
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目“MdmX促进非小细胞肺癌Mdm2抑制剂获得性耐药作用机理及其结构生物学机制研究”（32471260）;2024级校级大学生创新创业计划项目“老药新用-强心苷类化合物哇巴因抗结肠癌的作用机制研究”（XJU-SRT-24095）
- DOI：10.13568/j.cnki.651094.651316.2024.08.21.0001
  中图分类号： R735.35
- 纸质出版：2025
- 稿件说明：
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[1]徐建雄,努尔比亚木·达吾提,萨尼耶·依不拉音,等.哇巴因通过G6PD通路调节糖酵解影响结肠癌细胞增殖的机制研究[J].新疆大学学报(自然科学版中英文),2025,42(05):621-631.
[1]徐建雄,努尔比亚木·达吾提,萨尼耶·依不拉音,等.哇巴因通过G6PD通路调节糖酵解影响结肠癌细胞增殖的机制研究[J].新疆大学学报(自然科学版中英文),2025,42(05):621-631. DOI： 10.13568/j.cnki.651094.651316.2024.08.21.0001.

DOI：10.13568/j.cnki.651094.651316.2024.08.21.0001.

摘要

为探究哇巴因(ouabain)抑制结肠癌细胞HCT116增殖的作用机制，采用CCK-8法、细胞克隆形成实验、EdU实验和流式细胞术，分别检测了增殖能力、细胞克隆形成、DNA复制活性及凋亡率，以考察哇巴因对结肠癌细胞HCT116体外细胞活力的影响．为进一步分析哇巴因对结肠癌细胞HCT116糖酵解的影响，使用葡萄糖氧化酶法和乳酸检测试剂盒，检测了细胞葡萄糖消耗水平和乳酸生成水平的变化．此外，还探讨了哇巴因调控结肠癌细胞HCT116糖酵解的机制，通过real-time PCR分析其对相关代谢途径因子m RNA表达的影响，并通过Western Blot检测糖代谢相关蛋白G6PD的表达变化．结果表明：与对照组相比，哇巴因能显著抑制结肠癌细胞HCT116的增殖、克隆形成能力以及DNA复制活性(P<0.01)，其IC50值为50 nmol/L，并呈现浓度依赖性．同时，哇巴因能显著诱导结肠癌细胞HCT116凋亡(P<0.01)，并显著抑制结肠癌细胞HCT116的糖酵解，降低此代谢途径中相关因子mRNA的表达水平(P<0.01).Western Blot、realtime PCR和G6PD酶活力检测实验结果显示：与对照组相比，哇巴因可以显著降低结肠癌细胞HCT116中G6PD的mRNA和蛋白质表达水平以及G6PD酶活性(P<0.01)．综上所述，哇巴因可显著抑制结肠癌细胞HCT116的增殖，促进其凋亡并显著降低其糖酵解水平；其可能的机制与抑制G6PD信号通路有关，提示哇巴因可能成为治疗结肠癌的潜在候选药物，为结肠癌的治疗提供了新的思路和可能．

Abstract

To elucidate the mechanism of ouabain in the proliferation of colon cancer cells HCT116

the CCK-8assay

cell colony formation assay

EdU assay

and flow cytometry are employed to detect proliferation capacity

cell colony formation

DNA replication activity

and apoptosis rate

respectively

to investigate the effect of ouabain on the viability of colon cancer cells HCT116 in vitro.To further analyze the influence of ouabain on glycolysis in colon cancer cells HCT116

the glucose oxidase method and lactate detection kit are utilized to measure the changes in glucose consumption level and lactate production level of the cells.Additionally

the mechanism by which ouabain regulates glycolysis in colon cancer cells HCT116 is explored by analyzing the mRNA expression of related metabolic pathway factors via real-time PCR and detecting the expression changes of the glucose metabolismrelated protein G6PD through Western Blot.The experimental results reveal that

compared to the control group

ouabain significantly inhibits the proliferation

colony formation ability

and DNA replication activity of colon cancer cells HCT116(P<0.01)

with an IC50 value of 50 nmol/L

and exhibits a concentration-dependent manner.Meanwhile

ouabain also significantly induces apoptosis in colon cancer cells HCT116(P<0.01).Moreover

ouabain significantly suppresses glycolysis in colon cancer cells HCT116 and downregulates the mRNA levels of glycolytic metabolic pathway-related factors(P<0.01).The results obtained from Western Blot

real-time PCR

and cellular enzyme activity assays indicate that ouabain significantly reduces the mRNA and protein expression levels of G6PD

as well as decreases G6PD enzyme activity in colon cancer cells HCT116 compared to the control group(P<0.01).In conclusion

ouabain significantly inhibits the proliferation of colon cancer cells HCT116

promotes apoptosis

and suppresses glycolysis in these cells.The underlying mechanism appears to involve the inhibition of the G6PD signaling pathway.These findings suggest that ouabain holds potential as a candidate drug for colon cancer treatment

offering new avenues for therapeutic intervention in this disease.

关键词

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references

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