南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (2): 280-288.doi: 10.12122/j.issn.1673-4254.2024.02.10

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去甲泽拉木醛通过抑制 AKT/CREB 信号通路抑制非小细胞肺癌细胞的增殖、迁移和侵袭

韩齐齐，叶梦然，金齐力   

  1. 蚌埠医科大学检验医学院，安徽 蚌埠 233030；蚌埠医科大学第二附属医院检验科，安徽 蚌埠 233080
  • 发布日期:2024-03-13

Demethylzeylasteral inhibits proliferation, migration and invasion and promotes apoptosis of non-small cell lung cancer cells by inhibiting the AKT/CREB signaling pathway

HAN Qiqi, YE Mengran, JIN Qili   

  1. School of Laboratory Medicine, Bengbu Medical University, Bengbu 233030, China; Department of Laboratory Medicine, Second Affiliated Hospital of Bengbu Medical University, Bengbu 233080, China
  • Published:2024-03-13

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摘要： 目的 探讨去甲泽拉木醛抑制非小细胞肺癌细胞A549和H1299增殖、迁移和侵袭并诱导细胞凋亡的作用机制。方法 镜下初步观察不同浓度的去甲泽拉木醛（0、1、3、10、30 μmol/L）对A549和H1299细胞形态及细胞数量的影响。通过克隆形成、CCK-8、细胞划痕、Transwell和流式细胞实验检测去甲泽拉木醛对A549和H1299细胞增殖、细胞活力、细胞迁移和侵袭、细胞凋亡的影响以及SC79处理后对细胞凋亡的影响。通过Western blotting检测不同浓度的去甲泽拉木醛对A549和H1299细胞E-cadherin、N-cadherin、Vimentin、Bax、Bcl-2、cleaved-caspase 3表达和AKT/CREB磷酸化水平变化的影响，SC79处理后对凋亡蛋白表达的影响。结果 镜下观察药物作用后，细胞形态由饱满变为细长，细胞间隙增宽。CCK-8实验和克隆形成实验结果显示去甲泽拉木醛能抑制A549和H1299细胞活力并且抑制细胞的增殖（P<0.05）。流式细胞术结果表明，去甲泽拉木醛诱导A549和H1299细胞的凋亡，而SC79处理后逆转了促凋亡作用（P<0.05）。划痕和Transwell实验结果说明，去甲泽拉木醛抑制了A549和H1299细胞的迁移和侵袭（P<0.05）。Western blotting实验结果表明，去甲泽拉木醛处理后A549和H299细胞中Bax、clevaed-caspase3蛋白表达水平升高，Bcl-2蛋白表达水平下调，而SC79的使用逆转了凋亡蛋白的表达趋势，E-cadherin表达上调，N-cadherin、Vimentin表达下调，AKT/CREB磷酸化水平也被显著抑制（P<0.05）。结论 去甲泽拉木醛能够抑制非小细胞肺癌细胞的增殖、迁移和侵袭，并诱导其凋亡，其机制可能与其抑制AKT/CREB信号通路有关。

关键词: 非小细胞肺癌；去甲泽拉木醛；AKT；CREB

Abstract: Objective To investigate the mechanism underlying the inhibitory effects of Demethylzeylasteral (T-96) on non-small cell lung cancer (NSCLC) cells. Methods We first examined the effects of different concentrations (1, 3, 10, and 30 μmol/L) of demethylzeylasteral on morphology and cell number of A549 and H1299 cells. The changes in proliferation, cell viability, migration, invasion, and apoptosis of A549 and H1299 cells following demethylzeylasteral treatment were detected using clone formation, CCK-8, cell scratch, Transwell, and flow cytometric assays, and the effect of SC79 treatment against demethylzeylasteral-induced cell apoptosis was assessed. Western blotting was performed to detect the changes in expressions of E-cadherin, N-cadherin, vimentin, Bax, Bcl-2 and cleaved caspase-3 and phosphorylation of AKT/CREB in demethylzeylasteral-treated A549 and H1299 cells and the cellular expressions of apoptotic proteins following treatment with both demethylzeylasteral and SC79. Results T-96 treatment caused elongation of the cell body and widening of the intercellular space and significantly inhibited cell viability, proliferation, migration and invasion of A549 and H1299 cells (P<0.05). Flow cytometry showed that demethylzeylasteral induced apoptosis in both A549 and H1299 cells, whereas SC79 treatment obviously attenuated its pro-apoptotic effect (P<0.05). Western blotting revealed up-regulated expressions of Bax and cleaved caspase-3 proteins and lowered Bcl-2 expression level in demethylzeylasteral-treated A549 and H1299 cells, but co-treatment with SC79 obviously attenuated the expressions of the apoptotic proteins. T-96 significantly up-regulated the expression level of E-cadherin, down- regulated the expressions of N-cadherin and vimentin, and inhibited the phosphorylation of AKT and CREB in the two cell lines (P<0.05). Conclusion T-96 inhibits the proliferation, migration and invasion and induces apoptosis of NSCLC cells possibly by inhibiting the AKT/CREB signaling pathway.

Key words: non-small cell lung cancer; demethylzeylasteral; AKT; CREB