知母皂苷B-II通过Nrf2/miR-455-3p/KLF6途径抑制胃癌细胞增殖并诱导其凋亡。
Timosaponin B-II suppresses gastric cancer cell proliferation and induces apoptosis via the Nrf2/miR-455-3p/KLF6 pathway.
作者信息
Cui Qiaoyan, Zhang Ruijie, Zhao Shiqiao, Wang Jian, Dong Yafen, Bai Xuesong, Chen Ye, Qiu Yan
机构信息
Department of Pharmacy, Shanghai Pudong New Area People's Hospital, Shanghai, 201299, China.
Tongji University School of Medicine, Shanghai, China.
出版信息
Discov Oncol. 2025 May 23;16(1):894. doi: 10.1007/s12672-025-02650-9.
BACKGROUND
Gastric cancer (GC), known for its aggressive growth and metastasis, remains a leading cause of cancer-related mortality. Although Timosaponin B-II (TB-II) from Anemarrhena asphodeloides has shown anticancer potential, its underlying mechanisms in GC are not yet fully understood.
METHODS
Our study investigates and compares the expression patterns of Kruppel-Like Factor 6(KLF6) in GC tissues across different TNM stages, while exploring its upstream regulatory factors. GC cells were treated with Timosaponin B-II (TB-II), and alterations in cell proliferation and apoptosis rates were evaluated using CCK-8 and TUNEL assays. Furthermore, the expression levels of nuclear factor erythroid 2-related factor 2(Nrf2), miR-455-3p, and KLF6 were quantified to elucidate the mechanisms underlying TB-II's effects. To confirm the interaction between miR-455-3p and KLF6, as well as Nrf2 and miR-455-3p, bioinformatics analysis, luciferase assays, and ChIP-PCR were conducted. Finally, protein synthesis and degradation assays were performed to explore the mechanism by which TB-II regulates the expression and activity of Nrf2.
RESULTS
Both mRNA and protein expression levels of KLF6 were significantly lower in GC tissues compared to adjacent normal tissues. Notably, only KLF6 protein expression exhibited a decline in GC tissues from stages I to III of GC, whereas its upstream regulator, miR-455-3p, displayed the opposite trend. Treatment with TB-II markedly inhibited GC cells proliferation and induced apoptosis in a dose-dependent manner. Mechanistically, TB-II treatment upregulated the expression of Kelch-like ECH-associated protein 1(Keap1) protein, facilitating the formation of the Keap1/Nrf2 complex, which enhanced the ubiquitin-mediated degradation of Nrf2 in GC cells. Consequently, the transcriptional activation of miR-455-3p by Nrf2 was suppressed, resulting in the upregulation of the tumor suppressor KLF6. Silencing KLF6 can counteract the effects of TB-II in inhibiting proliferation and promoting apoptosis in GC cells.
CONCLUSION
TB-II suppresses GC cell proliferation and induces apoptosis via the Nrf2/miR-455-3p/KLF6 pathway.
背景
胃癌（GC）以其侵袭性生长和转移而闻名，仍然是癌症相关死亡的主要原因。虽然知母中的知母皂苷B-II（TB-II）已显示出抗癌潜力，但其在胃癌中的潜在机制尚未完全了解。
方法
我们的研究调查并比较了不同TNM分期的胃癌组织中 Kruppel样因子6（KLF6）的表达模式，同时探索其上游调节因子。用知母皂苷B-II（TB-II）处理胃癌细胞，使用CCK-8和TUNEL检测评估细胞增殖和凋亡率的变化。此外，对核因子红细胞2相关因子2（Nrf2）、miR-455-3p和KLF6的表达水平进行定量，以阐明TB-II作用的潜在机制。为了证实miR-455-3p与KLF6以及Nrf2与miR-455-3p之间的相互作用，进行了生物信息学分析、荧光素酶检测和染色质免疫沉淀PCR。最后，进行蛋白质合成和降解检测，以探索TB-II调节Nrf2表达和活性的机制。
结果
与相邻正常组织相比，胃癌组织中KLF6的mRNA和蛋白质表达水平均显著降低。值得注意的是，只有KLF6蛋白表达在胃癌I至III期组织中呈下降趋势，而其上游调节因子miR-455-3p则呈现相反趋势。TB-II处理以剂量依赖性方式显著抑制胃癌细胞增殖并诱导凋亡。机制上，TB-II处理上调了 Kelch样ECH相关蛋白1（Keap1）的表达，促进了Keap1/Nrf2复合物的形成，增强了胃癌细胞中Nrf2的泛素介导降解。因此，Nrf2对miR-455-3p的转录激活受到抑制，导致肿瘤抑制因子KLF6上调。沉默KLF6可抵消TB-II对胃癌细胞增殖的抑制作用和促进凋亡的作用。
结论
TB-II通过Nrf2/miR-455-3p/KLF6途径抑制胃癌细胞增殖并诱导凋亡。
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