Effects And Mechanism Of Melittin On The Proliferation Of Human Non-small Cell Lung Cancer A549 Cells

Objective: To investigate the effect of melittin on the proliferation of human non-small cell lung cancer A549 cells and its mechanism.Methods:(1)Human non-small cell lung cancer A549 cells were separately treated with melittin at 0 ng/ul,2.5 ng/ul,5 ng/ul,10 ng/ul and 20 ng/ul,the effect of melittin at different concentrations on the proliferation activity of human non-small cell lung cancer A549 cells was detected by CCK8 assay,and IC50 was calculated;(2)Human non-small cell lung cancer A549 cells were separately treated with 0ng/ul(control group)and 3ng/ul(MEL group)melittin,and the effect of melittin on the clone formation ability of human non-small cell lung cancer A549 cells was detected by cell clone formation assay;(3)The subcutaneous transplanted tumor of human non-small cell lung cancer A549 cells in nude mice was constructed.The nude mice injected with 100 ul melittin solution(dose of 5 mg/kg/mouse)intratumorally were used as the experimental group,and the nude mice injected with the same volume of normal saline intratumorally were used as the control group,changes in volume and weight of subcutaneously transplanted tumors in two groups of nude mice were detected;(4)Non-small cell lung cancer A549 cells were separately treated with 0ng/ul(control group)and 3ng/ul(MEL group)melittin,RNA-seq based on next-generation sequencing technology was used to analyze the effects of melittin on gene expression and signaling pathway in human non-small cell lung cancer A549 cells.Results:(1)Melittin could significantly inhibit the growth of human non-small cell lung cancer A549 cells in a dose-dependent manner within the concentration range of0-10ng/ul,and the inhibitory effect was significant in the concentration range of 10-20ng/ul.When the concentration reached 20ng/ul,almost all non-small cell lung cancer A549 cells died;(2)3 ng/ul Melittin could significantly inhibit the colony formation of human non-small cell lung cancer A549 cells(P<0.05).(3)Intratumoral injection of 100 ul melittin solution(dose of 5mg/kg/ mouse)could significantly inhibit the growth of human non-small cell lung cancer A549 cells subcutaneously transplanted in nude mice,and the inhibition rate was41.4% 14 days after administration(P <0.001).(4)A total of 1217 differentially expressed genes were found after RNA-seq.The differences in gene function were mainly reflected in the biological process of the angiogenesis and positive regulation of cell migration,and the differential gene expression in the PI3K-AKT signaling pathway was significant.Conclusion: Melittin may inhibit the proliferation of human non-small cell lung cancer A549 cells by regulating the expression of related genes in the PI3K-AKT signaling pathway.