L-securinine Induces Methylation Changes In The Human Lung Cancer Cell Line A549

Background: L-securinine is a class of natural alkaloids isolated and extracted from plants of the Euphorbiaceae family.Wnt antagonists consist of SFRP family and DKK family.Objective: To investigate the methylation changes of L-securinine in A549 cells and to explore its potential molecular mechanism as an antitumor agent.Methods: A549 cells were cultured with L-securinine.Cell counting kit-8(CCK-8)assay was used to assess the proliferation activity of A549 cells treated with L-securinine.Quantitative real-time PCR(q RT-PCR)and bisulfite sequencing PCR(BSP)were used to detect the expression and the DNA methylation of SFRPs genes and DKKs genes,respectively.Results: L-securinine inhibited the proliferation of A549 cells by dose-and time-dependent manner(P < 0.05).The IC50(half maximal inhibitory concentration)values at 24,36 and 48 h were 8.92,4.73 and 3.81?g/ml.The expression of SFRP1 gene in the A549 cells was lower than that in the HLF-a cells,DKK1 gene was in contrast.L-securinine induced the up-regulation of SFRP1 gene and down-regulation of DKK1 gene in the A549 cells and the methylation changes at Cp G sites in the SFRP1 and DKK1 promoter region,respectively.Conclusion: L-Securinine modulates the Wnt signaling pathway by inducing methylation changes of the SFRP1 gene and DKK1 gene.These observations indicate the potential of L-securinine as an antitumor active material targeting the Wnt signaling pathway.