Establishment Of Pancreatic Cancer Cell Lines Edited With KRAS Gene And Screening Of Potential Therapeutic Targets Based On Crispr/cas9 Technique

Objective: The purpose of this study was to establish a cell line with low expression of KRAS gene by CRISPR/Cas9,and to explore the relationship between KRAS gene and biological behavior of pancreatic cancer.The second is to use bioinformatics analysis to find out the gene F11 R which has the highest correlation with KRAS gene,and the third is to use RNA interference mediated by lentivirus vector to knock down the F11 R gene of pancreatic cancer cell line,and to explore the relationship between F11 R and biological behavior of pancreatic cancer.Looking for potential therapeutic targets for pancreatic cancer.Methods: The pancreatic cancer cell line with KRAS gene knockout was established by CRISPR/Cas9 and the cell line with low expression of KRAS gene was screened out.The related cell behavior experiments were carried out,including cell proliferation,cell scratch,cell invasion,cell cloning and so on.In addition,the proportion of apoptosis and the changes of cell cycle were analyzed by flow cytometry.Finally,the effect of the gene on the proliferation,migration and apoptosis of pancreatic cancer was analyzed statistically.F11 R knockout cell line was established on the basis of KRAS gene knock cell line of pancreatic adenocarcinoma,and the cell lines with low expression of F11 R gene were screened.The related cell behavior experiments were carried out,including cell proliferation,cell scratch,cell invasion,cell cloning and so on.Flow cytometry was used to analyze the percentage of apoptosis and the change of cell cycle.Finally,the data were analyzed to explore the effect of the gene on the proliferation,migration and apoptosis of pancreatic cancer.The low expression of F11 R gene in pancreatic cancer cell line was treated with antitumor drugs to determine whether the sensitivity of anticancer drugs to pancreatic cancer was changed and whether the gene was a potential therapeutic target for pancreatic cancer.Results:(1)KRAS gene knockdown pancreatic cancer cell line was established by CRISPR/Cas9,and B14 cell line with low expression of KRAS gene was screened out.(2)A series of biological behavior experiments demonstrated that knockout of KRAS gene tended to negatively regulate the malignancy of pancreatic cancer cells.(3)the highly correlated F11 R gene of KRAS gene was found by bioinformatics analysis,and the pancreatic cancer cell line of F11 R gene knockout was established by using lentivirus.(4)A series of biological behavior experiments confirmed that the malignant degree of pancreatic cancer cells was decreased after knockout of F11 R gene,and the sensitivity of anti-tumor drugs was increased.Conclusion:(1)after knockout the KRAS gene of pancreatic cancer,the malignancy of pancreatic cancer cells decreased.(2)as a highly correlated gene of KRAS gene,F11 R gene is involved in the proliferation,apoptosis,invasion and metastasis of pancreatic cancer cells,and affects the occurrence and development of pancreatic cancer.(3)after knockout of F11 R gene in pancreatic cancer cells,the sensitivity of anticancer drugs in pancreatic cancer cells increased,and F11 R gene may be a potential therapeutic target for pancreatic cancer.