Antitumor Activity Of A Novel Aurora A/B Kinases Inhibitor TY-011 Against Gastric Cancer By Inducing DNA Damage

Background The high incidence and high mortality rate of gastric cancer in China make it an important cancer type to treat.Traditional cytotoxic chemotherapeutic drugs,due to their lack of targeting and causing extensive damage to normal cells,have restricted their widespread use,resulting in occurrence of targeted tumor therapy.Aurora kinases are important kinases in mitosis and participate in important processes such as assembly and maturation of centrosomes,spindle formation,proper chromosome segregation,and cytoplasmic separation.In recent years,it has been found that Aurora kinase is highly expressed in a variety of tumors,including gastric cancer,and is involved in the occurrence,progression,and metastasis of tumors,making it an important target for tumor targeted therapy.As a carrier of intracellular genetic information,DNA is vulnerable to endogenous and exogenous factors that cause damage.DNA damage is an important cause of cell death.Aneuploidy caused by mitotic disorders is an important cause of DNA damage induction.In the process of cytokinesis that results in aneuploidy,the incorrectly-separated chromosomes are vulnerable to DNA double-strand breaks.The inhibition of Aurora kinase disturbs the correct separation of chromosomes and cytoplasm.Therefore,the relationship between Aurora kinase inhibition and DNA damage deserves further exploration.In the previous study,we screened a dual target compound TY-011 that targets both Aurora kinase and VEGFR,and found that it shows significant tumor suppression both in vivo and in vitro.It is a potential and highly effective oral administration,exhibiting low-toxic anti-tumor effect,owing further research value.Purpose To evaluate the effect of the novel Aurora kinase inhibitor TY-011 on the activity of Aurora kinase in gastric cancer cells and its effect on the proliferation of gastric cancer cells,and to evaluate the effect of this compound on DNA damage,in order to elucidate the specific mechanism of this compound and the relevance between Aurora kinase inhibition and DNA damage.Methods(1)Using Western blot to detect the change of the activities of Aurora kinase influenced by TY-011.(2)Using MTT assay to evaluate the anti-proliferation activity against several kinds of gastric cancer cells.(3)Using PI test to verify the influence on cell cycle made by TY-011.(4)Using Annexin V – FITC/PI method to test the influence on cell apoptosis made by TY-011.(5)Using alkaline comet assay to test the DNA damage induced by TY-011.(6)Using immunofluorescence and Western Blot to test the type of DNA damage induced by TY-011.Results(1)TY-011 inhibits Aurora A/B kinases activities in vitro.(2)TY-011 shows in vitro antitumor effects in a variety of gastric cancer cells.(3)TY-011 induces cell cycle arrest and polyploidy(4)TY-011 induces apoptosis(5)TY-011 induces DNA damage(6)TY-011 induces DNA double strand breaks(DSBs)indicated by activation of DNA damage response molecules.Conclusion Compound TY-011 can significantly inhibit the activity of Aurora kinase in gastric cancer cells,effectively arrest gastric cancer cells in G2/M phase and induce apoptosis,and can also induce DNA damage in gastric cancer cells,and ultimately inhibit the proliferation of gastric cancer cells.It provides a new mechanism for TY-011 to play a role in anti-gastric cancer,elucidates the relationship between Aurora kinase inhibition and DNA damage,provides experimental basis for DNA damage induced by Aurora kinase inhibitors,and provide theoretical basis and design ideas for novel tumor targeted therapeutic drug R & D and mechanism research.