Effects Of RNase A On Biological Behavior And Functions Of A549 Cells In Vitro

Objective: Tumors are a major human disease.Althrough the traditional surgical resection,chemotherapy,radiation therapy,and immunotherapy have their advantages,they also have several limitations in clinical application.Therefore,it is necessary to develop new antitumor drugs with specificity and efficacy.RNase,as a kind of widespread natural enzymes with cytotoxicity,has gradually been proven to have potential as an antitumor drug.Studies have shown that different RNase have multiple effects on cancer under different conditions,but its specific functional mechanism remains to be further elucidated.To this end,the human non–small-cell lung cancer A549 cells as a model was used in this study.We have explored the effects of RNase A on biological behaviors and functions of A549 cells,and further revealed its mechanisms of functions.The research results will provide a theoretical basis for the development of RNase-based antitumor Drugs.In addion,we have preliminary studied the effects of RNase A on the proliferation of E.coli DH5? in order to provide the evidence for further explaining the relationship between RNase A and bacterial infection.Methods: Part one.The effects of RNase A on the biological behavior and function of A549 cells.1)The effects of RNase A on the morphology of A549 cells were observed by optical microscope and laser confocal scanning microscope,respectively.2)Transwell assay was used to detect the effects of different concentrations of RNase A on migration and invasion of A549 cells.3)Sphere-formation assay was used to measure the ability of sphere formation of different concentrations of RNase A on A549 cells.4)CCK-8 assay was used to detect the effects of different concentrations of RNase A on proliferation of A549 cells.5)CCK-8 assay was used to detect the effects of different concentrations of RNase A combined with adriamycin and cisplatin on A549 cells.Part two.The effects of RNase A on the proliferation of E.coli DH5?.1)E.coli DH5? strains were cultured with different concentrations of RNase A in LB or M9 media,and the growth curves were plotted to detect effect of RNase A on the proliferation of E.coli DH5?.2)The effect of RNase A on the proliferation of E.coli DH5? was detected in LB medium pretreated with RNase A.3)Colony formation assay was used to detect the effect of RNase A on the colony formation of E.coli DH5?.Results: Part one.The effects of RNase A on biological behavior and function of A549 cells.1)RNase A caused scatter response of A549 cells and induced numerous pseudopodia on the surface of A549 cells.2)Low and medium concentrations of RNase A significantly promote migration and invasion of A549 cells.3)RNase A inhibits sphere-formation in A549 cells.4)Low concentration of RNase A promotes the proliferation of A549 cells,and high concentration of RNase A inhibits the proliferation..5)RNase A combined with adriamycin and cisplatin has a cumulative cytotoxic effect on A549 cells.Part two.The effects of RNase A on the proliferation of E.coli DH5?.1)Different concentrations of RNase A promote the proliferation of E.coli DH5? in LB and M9 medium.2)RNase A promotes colony formation of E.coli DH5?.Conclusion: RNase A affects a variety of biological behaviors and functions of A549 cells in vitro.These phenomena might be caused by functional diversity of RNase A,and its functional mechanism needs to be further studied.In addition,RNase A promotes the proliferation of E.coli DH5? in LB and M9,and its mechanism needs to be further elucidated.