去甲基泽拉斯特醇通过抑制AKT/CREB信号通路抑制非小细胞肺癌细胞的增殖、迁移和侵袭并促进其凋亡
[Demethylzeylasteral inhibits proliferation, migration and invasion and promotes apoptosis of non-small cell lung cancer cells by inhibiting the AKT/CREB signaling pathway].
作者信息
Han Q, Ye M, Jin Q
机构信息
School of Laboratory Medicine, Bengbu Medical University, Bengbu 233030, China.
Department of Laboratory Medicine, Second Affiliated Hospital of Bengbu Medical University, Bengbu 233080, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Feb 20;44(2):280-288. doi: 10.12122/j.issn.1673-4254.2024.02.10.
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 ( < 0.05). Flow cytometry showed that demethylzeylasteral induced apoptosis in both A549 and H1299 cells, whereas SC79 treatment obviously attenuated its pro-apoptotic effect ( < 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 cotreatment 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 ( < 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.
目的
探讨去甲基泽拉斯醇（T-96）对非小细胞肺癌（NSCLC）细胞抑制作用的潜在机制。
方法
我们首先检测了不同浓度（1、3、10和30 μmol/L）的去甲基泽拉斯醇对A549和H1299细胞形态和细胞数量的影响。使用克隆形成、CCK-8、细胞划痕、Transwell和流式细胞术检测去甲基泽拉斯醇处理后A549和H1299细胞增殖、细胞活力、迁移、侵袭和凋亡的变化，并评估SC79处理对去甲基泽拉斯醇诱导细胞凋亡的影响。进行蛋白质免疫印迹法检测去甲基泽拉斯醇处理的A549和H1299细胞中E-钙黏蛋白、N-钙黏蛋白、波形蛋白、Bax、Bcl-2和裂解的半胱天冬酶-3表达的变化以及AKT/CREB的磷酸化情况，以及去甲基泽拉斯醇和SC79联合处理后细胞凋亡蛋白的表达情况。
结果
T-96处理导致细胞体伸长和细胞间空间增宽，并显著抑制A549和H1299细胞的细胞活力、增殖、迁移和侵袭（<0.05）。流式细胞术显示去甲基泽拉斯醇诱导A549和H1299细胞凋亡，而SC79处理明显减弱其促凋亡作用（<0.05）。蛋白质免疫印迹法显示，去甲基泽拉斯醇处理的A549和H1299细胞中Bax和裂解的半胱天冬酶-3蛋白表达上调，Bcl-2表达水平降低，但与SC79联合处理明显减弱了凋亡蛋白的表达。T-96显著上调两种细胞系中E-钙黏蛋白的表达水平，下调N-钙黏蛋白和波形蛋白的表达，并抑制AKT和CREB的磷酸化（<0.05）。
结论
T-96可能通过抑制AKT/CREB信号通路抑制NSCLC细胞的增殖、迁移和侵袭并诱导其凋亡。
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