The Anti-tumor Effect Of Aurora-A Kinase Inhibitor SPAUFK9 On Gastric Cancer Cells

Gastric cancer is one of the most widespread malignant digestive tract cancers in the world,with prominent morbidity and mortality.Aurora-A kinase can regulate cell mitosis and act as an oncogene in many solid tumors.SPAUFK9 is a molecular targeted drug and a specific inhibitor of Aurora-A kinase.In vitro experiments,SPAUFK9 has shown anticancer effects in breast and lung cancer,However,its anti-tumor effects on gastric cancer still unclear.Objective: To study the expression and clinical significance of Aurora-A kinase in gastric cancer tissues and explore the anticancer effect of Aurora-A kinase inhibitor SPAUFK9 on gastric cancer cells,and provide more evidence for the molecular targeted therapy of gastric cancer.Methods:1.We combine TCGA database and GTEx database to search and detect fresh tissue samples of gastric cancer patients by WB experiment to analyze the expression of Aurora-A;2.A retrospective collection of 86 gastric cancer patients admitted to the Department of General Surgery of the 940 Hospital of the Joint Logistics Service Force from March 2014 to January 2015.We collected the wax blocks of cancer tissue and normal tissue wax blocks,We detected the expression level of Aurora-A by Immunohistochemical staining method and analyzed the relationship between the expression level of Aurora-A and the clinical pathology of patients.3.We used CCK-8 experiment to explore the effect of SPAUFK9 on the proliferation of gastric cancer cells;We used the scratch experiment and Transwell experiment to detect the effect of SPAUFK9 on gastric cancer cell migration and invasion.We used the flow cytometry to observe the influence of SPAUFK9 on gastric cancer cell cycle and apoptosis.We used the RT-PCR experiment to detect the changes in the expression level of Aurora-A m RNA in gastric cancer cells;We used WB assay to detect the changes in the expression levels of Aurora-A protein,apoptosis-related proteins Bax and Bcl-2;the key protein Yap of the Hippo signaling pathway and P21 protein.Results:1.Through a joint search of the TCGA database and the GTEx database,We obtained RNA sequencing data of 352 gastric cancer samples and 192 normal human gastric samples,and analyzed the differences in the expression of Aurora-A m RNA.The results showed that Aurora-A expression in gastric cancer tissues was higher than normal gastric tissues.At the same time,we extracted the total protein from fresh gastric cancer tissues and normal gastric tissues from 10 patients with gastric tumor.By WB test,we found that the expression of Aurora-A protein in cancer tissues was higher than that in normal gastric tissues.2.We selected 86 cases of gastric cancer patients with cancer tissue and normal gastric tissue wax blocks,applied immunohistochemical staining method to detect the relationship between the expression level of Aurora-A and the patient’s clinical pathology.The results showed that compared with normal tissues adjacent to cancer,Aurora-A was overexpressed in gastric cancer tissues,and was related to tumor pathological type,degree of differentiation,tumor maximum diameter,depth of invasion,TNM stage,lymph node metastasis,vascular invasion,and nerve invasion(P<0.05);there was no obvious correlation with age,gender,and tumor location(P>0.05);Multivariate Cox regression analysis showed that vascular invasion and Aurora-A protein expression can be used as independent related risk factors for the overall survival of gastric cancer patients;Kaplan-Meier survival analysis results showed that gastric cancer patients with low expression of Aurora-A protein had a longer overall survival time than gastric cancer patients with overexpression of Aurora-A protein(P<0.05).3.The Aurora-A kinase inhibitor SPAUFK9 inhibited the proliferation,migration and invasion of gastric cancer cells,causing gastric cancer cells to exhibit G2/M phase block,inducing an increase in the total apoptosis rate of gastric cancer cells(P<0.05);Aurora-A kinase inhibitor SPAUFK9 decreased the expression of Aurora-A m RNA,Aurora-A protein,Bcl-2 protein and Yap protein,and increased the expression of Bax and P21 protein at the molecular level(P < 0.05).Conclusions:1.Aurora-A is overexpressed in gastric cancer tissues and is closely related to the depth of tumor invasion,TNM stage,degree of differentiation,lymph node metastasis,and vascular invasion,and it is significantly related to the poor prognosis of gastric cancer patients.2.SPAUFK9 can inhibit the proliferation,migration and invasion of gastric cancer cells,block the cycle process of gastric cancer cells,and promote apoptosis of gastric cancer cells.3.SPAUFK9 effectively acts on the Aurora-A target of gastric cancer cells,the mechanism of influence on the apoptosis of gastric cancer cells may be related to "SPAUFK9-Aurora-A kinase-Yap/P21".