| Background Gastric Cancer(GC)is the fourth most familiar malignant neoplasm worldwide,and there has been no significant progress in survival in recent years,and the survival rate after 5 years is still less than 30%.Targeted CDK4/6 therapy is the current hot spot in the development of anti-tumor drugs,in 2020 CDK4/6 inhibitor abecept was approved by the state,combined endocrine therapy for HR+,HER2-advanced or metastatic breast cancer adjuvant therapy,but CDK4/6 inhibitors in human gastric cancer cells are less studied,the role is not clear,cisplatin as a common chemotherapy drug in gastric cancer,side effects are obvious,easy to produce drug resistance.Therefore,studying the effect of CDK4/6inhibitors on the biological behavior of gastric cancer cells will be of great significance for the treatment of gastric cancer.Objective The role of CDK4/6 inhibitors in human gastric cancer cells is unclear,and the side effects of cisplatin are obvious,this study uses CDK4/6 inhibitors and cisplatin in combination with human gastric cancer cells to reduce the amount of cisplatin and achieve the same efficacy,and to investigate the impact on different biological behaviors of human stomach cancer cells.Methods 1.It is conducive to the online analysis of TCGA database resources on the bioinformatics UALCAN website(http://ualcan.path.uab.edu/)to obtain the expression of CDK4 and CDK6 genes in gastric cancer and normal gastric tissues;2.Western blot test was used to detect the representation of CDK4 and CDK6 albumen in stomach carcinoma cell line SGC7901 cells and regular gastric mucous membrane cell line GES-1 cells;3.CCK8 method to detect the OD values of distinct density of cisplatin(0μg/ml,0.25μg/ml,0.5μg/ml,1μg/ml,2μg/ml)and abemaciclib(0μM,5μM,10μM,20μM,40μM),and screen the optimal cell growth inhibition concentration of cells,that is,half of the inhibitory concentration(IC50);4.According to the optimal cell growth inhibition concentration obtained by the above experiments,the experiments were divided into groups,and the impact of the associated of abemaciclib and cisplatin on cell multiplication capacity was debated;5.According to the above experimental grouping,the apoptosis standard of every set of cells was measuring by Annexin v-FITC/PI dual tinged streaming,and the impact of the united action of abemaciclib and cisplatin on apoptosis was explored;6.The relocation capacity of every set of cells was measuring by scratch relocation laboratory,and the impact of the combination of albecillib and cisplatin on cell migration ability was discussed;7.The invasion competence of every set of cells was examine by Transwell laboratory,and the effect of the combination of abemaciclib and cisplatin on cell invasion ability was explored.Results1.Analysis of TCGA database resources through the bioinformatics UALCAN website,It was discovered that the representation of CDK4 and CDK6 albumen in stomach cancer cell microstructure was obvious taller than that in regular stomach microstructure(P<0.05);2.Compared with GES-1 cells of normal stomach mucous membrane cell line,the representation of CDK4 protein and CDK6 protein in SGC7901 cells was evident rised(P<0.05);3.After the analysis of CCK-8 experimental results,the cisplatin IC50 value was(0.5μg/ml)and the abemaciclib IC50 value was 10(μM).4.Carry out subsequent experiments based on the optimal concentration of the drug obtained from the above experiments,and divide the experiment into a blank control group,cisplatin group,cisplatin+abemaciclib group,abemaciclib group,the inhibition rate can be calculated through the CCK-8 laboratory,compared to the blank control group,cisplatin group and abemaciclib group,the restrain rate of cisplatin + abemaciclib group was significantly elevated(P<0.01),and the restrain capacity was more evident with the prolongation of the action time;5.The apoptosis rate of cells in each group was inspection by flow apoptosis laboratory,and compared with the blank control set,the cisplatin set and the abemaciclib set could remarkable strengthen the apoptosis proficiency of cells(P<0.01).6.After the analysis of scratch experiment results,compared with the blank control group,cisplatin group and abemaciclib group,the cell migration capacity of the cisplatin + abemaciclib group was significantly reduced(P<0.01);7.The invasion capacity was examine by the Transwell laboratory,compared with the blank control group,cisplatin group and abemaciclib group,cisplatin + the cell invasion ability of the abemaciclib group was significantly weakened(P<0.01).Conclusion This study found that the representation of CDK4 and CDK6 in stomach cancer tissues and human gastric cancer cells was remarkable elevated.Compared with the drug alone,the CDK4/6 inhibitors abemaciclib and cisplatin showed a synergistic effect,which can significantly restrain the proliferative ability of human stomach cancer cells,promote apoptosis,and weaken their migration and invasion ability,indicating that the combination of abemaciclib and cisplatin may achieve better therapeutic effects than monotherapy in the treatment of gastric cancer,providing a new idea for the treatment of gastric cancer. |
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