| Objective: Gastric carcinoma(GC)is one of the most common malignant tumors,and the morbidity and mortality of gastric carcinoma ranks third in China.Nowadays,individualized treatment based on the combination of surgery,radiotherapy and chemotherapy is adopted in our country,but because most of patients are found to be in advanced stage,the prognosis of gastric carcinoma has not been substantially improved.Previous studies have shown that the occurrence and development of GC are complex process involving multi-genes,multi-steps and multi-factors.It is necessary to further explore the development and the possible molecular mechanisms of gastric carcinoma to provide new biomarkers and therapeutic targets.Eukaryotic translation initiation factor 6(e IF6)is one of the eukaryotic translation factors,which plays an anti-binding role in the process of translation initiation and participates in the process of ribosome formation.In recent years,studies have shown that e IF6 is overexpressed in a variety of malignant tumors,which is likely to be an oncogene,but the role and mechanism of e IF6 in gastric carcinoma is not clear.The purpose of this study is to explore the role and function of e IF6 in gastric carcinoma so as to provide a new theoretical basis for the development of gastric carcinoma.Methods:1.Immunohistochemical technique was used to detect the expression of e IF6 in gastric carcinoma tissues,and the relationship between the expression of e IF6 in cancer tissues and clinicopathological characteristics were analyzed.2.AGS and MGC803 with high e IF6 m RNA level and protein expression were screened from the gastric carcinoma cell lines HGC27,SGC7901,AGS,MGC803 and BGC823 cell lines by RT-q PCR test and Western Blot test.3.si RNA-2 and si RNA-3 with high silencing efficiency were screened from three si RNA fragments of e IF6 gene by RT-q PCR and Western Blot experiments.4.The e IF6 gene was silenced in AGS and MGC803 cell lines by si RNA transfection technique.CCK8 test,plate cloning test,scratch test and Transwell invasion test were used to detect the rols of e IF6 on the proliferation,colony cloning,migration and invasion of AGS and MGC803 cells in vitro.5.Flow cytometry was used to detect the rols of e IF6 on cell cycle of AGS and MGC803.Western blot assay was used to detect the effect of e IF6 on the expression of cyclin B1 in AGS and MGC803.Results:1.Immunohistochemical results showed that the expression of e IF6 in cancer tissues was significantly higher than that in adjacent tissues(P<0.001).The expression of e IF6 was significantly correlated with p TNM stage(P<0.007),but not significantly correlated with age,gender,tumor size,degree of differentiation,lymph node metastasis and distant metastasis(P>0.05).2.The results of CCK8 test showed that the OD value of AGS/MGC803 cell line after e IF6 silencing was significantly lower than that of the control group at24 h,48h,72 h and 96 h,which indicated that e IF6 silencing could reduce the proliferation of gastric carcinoma cells.3.The results of plate cloning experiment showed that the colony formation number of AGS/MGC803 cell line after silencing e IF6 was significantly reduced compared with the control group,which indicated that silencing e IF6 could reduce the colony formation ability of gastric carcinoma cells.4.The results of scratch test showed that the migration rate of AGS/MGC803 cell line after silencing e IF6 was lower than that of the control group at 24 h and 48 h,which indicated that silencing e IF6 could reduce the migration ability of gastric carcinoma cells.5.The results of the Transwell invasion experiment showed that the AGS/MGC803 cell line after silencing e IF6 was significantly reduced compared with the cells that passed through the compartment in the control group,indicating that silencing e IF6 can reduce the invasion ability of gastric carcinoma cells.6.Flow cytometry was used to detect the cell cycle of AGS/MGC803 cell line after e IF6 silencing,and the results showed that e IF6 silencing blocked gastric carcinoma cells in the G2/M phase,and Cyclin B1 protein level was reduced.Conclusion:1.The expression of e IF6 in gastric carcinoma tissues were significantly higher than that in adjacent tissues,and its positive intensity was related to p TNM.2.Silencing e IF6 reduced the ability of proliferation,colony formation,migration and invasion of AGS and MGC803 cells.It is speculated that e IF6 plays a cancer-promoting role in gastric carcinoma.3.Silencing e IF6 could block the cell cycle of gastric carcinoma cells AGS and MGC803 in G2/M phase and decrease the expression of Cyclin B1. |
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