The Study Of Knockout ABCG2 By CRISPR-Cas9 System On The Malignancy Of Colorectal Cancer Cell S1-M1-80

Background: CRISPR(Clustered regularly interspersed short palindromic repeats)can resist the invasion of virus and exogenous DNA.CRISPR-Cas9 system can utilize sgRNA to lead Cas9 endonuclease to cut the genomic double-strand DNA in order to achieve the effect of gene knockout.Overexpressed ABCG2 protein in tumor cell lead to generate multidrug resistance and other malignant behavior which will cause the failure of therapy.In this study we used CRISPR-Cas9 system to knockout ABCG2 to study the function of ABCG2 in colorectal cancer.Method: Lenti CRISPR v2 vectors were constructed.Lentivirus infection was performed to construct ABCG2 knockout cell.MTT assay was used to investigate the cell growth rate and multidrug resistance.Drug accumulation was performed to study the ability of drug accumulation.Flow cytometry was applied to study cell cycle.Wound healing,Transwell and Soft agar were used to check the ability of cell migration,invasion and colony formation.Result: ABCG2 gene knockout vector and stable cells line were constructed success.The drug sensitivity of ABCG2 gene knockout cell line was increased.Drug resistance of ABCG2 gene knockout cell line was reduced.Cell growth rate of ABCG2 gene knockout cell line was not changed but cell cycle was changed.The abilities of migration,invasion and colony-formation after knockout ABCG2 in colorectal cancer were all inhibited.Conclusion: These data demonstrate that knockout ABCG2 can inhibit the malignancy in colorectal cancer by CRISPR-Cas9 system...