Found programs:
Authors:Liu Ping; Geng Yadi; Liu Yunxiao; Xie Yanbo; Zhang Xinge; Zhang Lei; Wei Wei
Keywords:apigenin;NCI-H520;proliferation;metastasis;apoptosis;autophagy
DOI:10.19405/j.cnki.issn1000-1492.2022.07.001
〔Abstract〕 Objective To investigate the effect of apigenin on the proliferation, migration, invasion, apoptosis and autophagy of human lung squamous carcinoma NCI-H520 cells. Methods Human lung squamous carcinoma NCI-H520 cells were culturedin vitro, and the CCK-8 method was used to detect the effects of different concentrations of apigenin(2.5, 5, 10, 20 μmol/L) or cisplatin(2.5, 5, 10, 20 μmol/L) on cell viability. Carboxyfluorescein diacetate, succinimidyl ester(CFDA SE) was used to detect the effect of apigenin or cisplatin on cell division. Scratch test and Transwell test were used to detect the effect of apigenin on cell migration and invasion. Annexin V/PI double staining method and Hoechst 33258 nuclear staining method were used to detect the effect of apigenin on cell apoptosis. Transmission electron microscopy was used to observe the generation of autophagic vesicles in cells. Acridine orange(AO) staining was used to observe the changes of acidic organelles in cells. Western blot was used to detect microtubule-associated protein 1 light chain 3 B-Ⅱ(LC3 B-Ⅱ) and p62 protein expression. Results Compared with the control group, CCK-8 assay and CFDA SE showed that apigenin or cisplatin reduced proliferation of NCI-H520 cells(P<0.05). Scratch test and Transwell test showed that apigenin reduced the migration and invasion levels of cells(P<0.01). Annexin V/PI double staining showed that apigenin increased apoptosis rate(P<0.05). Hoechst 33258 nuclear staining showed that apigenin promoted nuclear condensation and hyperchromatism of cells. AO staining fluorescence value enhanced, the autophagy bimolecular structure appeared under transmission electron microscopy, and the results of LC3 B-Ⅱ and p62 protein expression levels in Western blot showed that apigenin increased the autophagy level of cell NCI-H520(P<0.05). Chloroquine(CQ) failed to increase the protein levels of LC3 B-Ⅱ and p62 in apigenin treated cells. Conclusion Apigenin at different concentrations can inhibit the proliferation, migration and invasion of human lung squamous cell carcinoma NCI-H520, and its inhibition of cell growth and metastasis may be related to the induction of apoptosis and autophagy, and the increase of autophagy may be caused by blocking autophagosome degradation.