Study On The Role And Mechanism Of MiRNA-584-5p Targeted Regulation Of LRRC19 In The Proliferation,Invasion,Migration And Apoptosis Of Colorectal Cancer Cells

Objective: According to the statistics of the Global Cancer Center in 2020,colorectal cancer have become the third largest malignant tumors in the world,with a mortality rate second only to lung cancer,and it is increasingly showing a younger trend.Micro RNA(miRNA)is an endogenously expressed micro-noncoding RNA composed of 20-24 nucleotides that regulates cellular processes such as cell proliferation,apoptosis,migration,invasion,and differentiation,and passes through the m RNA of the target gene3,UTR-end binding inhibits the translational regulatory function of its target gene and plays a role.Abnormal expression of miRNA has also been reported in many malignancies.Many studies have shown that miRNA can play a role as a specific target in the diagnosis and treatment of malignant tumors.miRNA-584-5phas been shown to play an important regulatory role in various malignancies such as gastric cancer,lung cancer,hepatocellular carcinoma,breast cancer,etc.Studies have confirmed that miRNA-584-5pis highly expressed in the tissues and plasma of colorectal cancer patients,but how it plays a role in colorectal cancer and its target genes have not been clearly studied.The purpose of this study is to explore the regulatory role and mechanism of miRNA-584-5 p and its target gene in colorectal malignancies,so as to find new markers and therapeutic targets for the diagnosis and treatment of colorectal malignant tumors.Methods:(1)Colorectal cancer cells(SW480,SW620,HT-29,DLD-1)and colonic normal epithelial cells were detected by applying real-time quantitative polymerase chain reaction q RT-PCR The relative expression of miRNA-584-5P in the line NCM460 cells was statistically analyzed for expression differences.(2)After transfecting SW480 colorectal cancer cell line with miRNA-584-5p inhibitor and miRNA-584-5p-NC,perform CCK8,Transwell,cell scratch experiment,cell cycle detection,A series of in vitro functional experiments such as apoptosis detection to observe the proliferation,migration and invasion ability of cells during miRNA-584-5p silencing,and whether the cycle and apoptosis of cells are affected.(3)The target gene of m-iRNA-584-5p was analyzed by bioinformatics software,and the relationship between miRNA-584-5p and the target gene was verified by diluciferase experiment.(4)Colorectal cancer cells(SW480,SW620,HT-29,DLD-1)and colonic normal epithelial cells were detected by applying real-time quantitative PCR(Real-time quantitative polymerase chain reaction q RT-PCR)method Relative expression of m RNA of the target gene in the NCM460 cells and using western The blot experiment detects the relative expression of target gene proteins in each cell line,and finally uses statistics to analyze whether there are differences in expression.(5)A series of in vitro functional experiments such as CCK8,Transwell,cell scratch experiment,cell cycle detection,apoptosis detection were carried out after upregulating the target gene or simultaneously upregulating the target gene and m-iRNA-584-5p,which further showed that m-iRNA-584-5p acted on the target gene to regulate cell function.Results:(1)q RT-PCR results showed that the expression of miRNA-584-5p in colorectal cancer cell lines was higher than that of normal colon cancer epithelial cells.(2)CCK8,Transwell,cell scratch experiments showed that when miRNA-584-5p was knocked down,the proliferation ability,migration ability and invasion ability of colorectal cancer cells were inhibited,that is,miRNA-584-5p played the role of oncogene in colorectal cancer.(3)The bioinformatics method predicted that LRRC19 was the target gene of miRNA-584-5p,and it was confirmed by diluciferase experiments.(4)The results of q RT-PCR and western blot experimental showed that the expression of LRRC19 in colorectal cancer cell lines was lower than that of normal colon cancer epithelial cells.(5)When the expression quantity of LRRC19 was upregulated,the proliferation ability,migration ability and invasion ability of colorectal cancer cells decreased,the ratio of G2/M phase in the proportion increased,and the proportion of apoptosis also increased.After upregulating expression quantity of LRRC19 and miRNA-584-5p in colorectal cancer cells,the proliferation capacity,migration capacity and invasion capacity of colorectal cancer cells increased compared with LRRC19 overexpression,and the ratio of G2/M phase to apoptosis decreased compared with LRRC19.Conclusions:(1)miRNA-584-5p was highly expressed in colorectal cancer cell lines and low expression in normal epithelial cell lines in the colon;(2)miRNA-584-5p has a targeted regulatory relationship with LRRC19;(3)In the pathological development of colorectal cancer,miRNA-584-5p reflects carcinogenicity,LRRC19 reflects tumor suppression: miRNA-584-5p promotes the proliferation and invasion of colonic malignant tumor cells and inhibits apoptosis by negatively acting on LRRC19.