Function Study On The SCRC Candidate Gene DDI2,Based On Whole Exome Sequencing Technology

In recent years,with the rapid development of China’s economy and changes in living habits,the incidence and mortality of sporadic colorectal cancer are also rising,and sCRC has become a kind of malignant tumor that seriously endangers human health.According to the domestic and foreign research,it is found that the changes of KRAS,P16 and TP53 are closely related to the occurrence of colorectal cancer,but 30%of colorectal cancer is not caused by this mechanism.Therefore,it is great significance to explore the new mechanism and the candidate genes of colorectal cancer.Whole Exome Sequencing Technology is a highly sensitive method for genomic analysis that can find a large number of disease-related mutations by sequencing 1%of the genome.Therefore,this study uses Whole Exome Sequencing Technology to identify the possible candidate gene DDI2 of sporadic colorectal cancer,and LoVo cells were selected to carry out gene overexpression function experiments,which laid the foundation for the follow-up study.MethodsFirst,Whole Exome Sequencing Technology was used to screen out the loss of function mutations in three samples of sCRC cancer tissue,and the candidate gene DDI2 was identified by the correlation bioinformatics analysis method.Then,according to the results of previous studies,the colorectal cancer cells LoVo,which had the lowest gene expression level,was selected and transfected with plasmid pIRES2-EGFP,pIRES2-DDI2 and pIRES2-DDI2 mutant respectively.Some experiments In vitro were performed to observe the effect of DDI2 overexpression on the biological characteristics of CRC cells.Result1)Through the scratch experiment,Transwell migration experiment,Transwell invasion experiment,CCK-8 proliferation experiment and clonal formation experiment,We found compared the control group with DDI2 wild group,the migration,invasion and proliferation ability of experimental group cell were weakened,and the difference was statistically significant.2)The apoptosis of the cells was detected by flow cytometry.Compared with the control group,the experimental group found that there was no significant difference in the apoptosis rate between the two groups,which indicated that the effect of DDI2 on the proliferation of LoVo cells might not be caused by apoptosis.3)Cell viability at different concentrations of 5-FU was measured by CCK-8,we found that the survival rate of experimental group was lower than that of control group,and there was significant difference in 5-FU concentration.The results also showed that DDI2 overexpression could decrease the survival rate of LoVo cells,Of the drug resistance.CONCLUSIONDDI2 overexpression can reduce the proliferation,migration and invasion of LoVo cells and reduce the resistance of LoVo cells to 5-FU.It also suggests that DDI2 may be involved in the molecular mechanism of sCRC.