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、30μmol/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。
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 30μmol/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 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(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.
作者
韩齐齐
叶梦然
金齐力
HAN Qiqi;YE Mengran;JIN Qili(School of Laboratory Medicine,Bengbu Medical University,Bengbu 233030,China;Department of Laboratory Medicine,Second Affiliated Hospital of Bengbu Medical University,Bengbu 233080,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2024年第2期280-288,共9页
Journal of Southern Medical University
基金
安徽省高校自然科学研究重点项目(2023AH051974)。
