Effects Of MiR-203a-3p On The Proliferation And Migration Of Pancreatic Carcinoma Cells

Objectives The experiment examined the effect of miR-203a-3p on the proliferation,migration and invasion ability of pancreatic cancer cell lines and provided an experimental basis for studying the invasion and metastasis mechanism of pancreatic cancer and specific therapeutic targets.Methods 1 Screen The Cancer Genome Atlas(TCGA)database for differential expression of miRNAs in pancreatic cancer tissues,analyze heat maps;select miRNAs with species conservation and use Kaplan-Meier to analyze the overall survival rate of miRNAs in low and high-expression groups of pancreatic cancer tissues and draw survival curves and clinical staging maps,screened for miRNAs with statistical significance: miR-192-5p,miR-203a-3p,miR-451 a.2 Analysis of cancer-related GO and KEGG enrichment pathways using the Tarbase database.3 In pancreatic cancer cell lines Bxpc-3,Aspc-1,Panc-1 and human normal pancreatic epithelial cells HPNE,the expression levels of miR-192-5p,miR-203a-3p and miR-451 a were verified using real-time quantitative PCR technology(hereinafter referred to as qRT-PCR),miR-203a-3p was selected as the research object.4 CCK-8 test to determine the effect of miR-203a-3p on the proliferation of pancreatic cancer cell lines Bxpc-3 and Panc-1.5 Transwell experiments detects the effects of miR-203a-3p on the migration and invasion of pancreatic cancer cell lines Bxpc-3 and Panc-1.6 Colony formation experiments to determine the effect of miR-203a-3p on the colony forming ability of pancreatic cancer cell lines Bxpc-3 and Panc-1.7 Apply DIANA TOOLS,Mi RDB and Target Scan to predict the target genes of miR-203a-3p,analyze the function of PPM1 A genes and make a gene interaction network map.Results 1 A total of 18 miRNAs with a differential expression greater than 2 times normal pancreatic tissue and tumor tissue were selected through the TCGA database.Among them,misas with species conservation were miR-192-5p,miR-203a-3p,miR-451a;the KaplanMeier curve,clinical cancer stage,enrichment function and expression pathway of three miRNAs were statistically significant(P<0.05).2 qRT-PCR results showed that miR-192-5p and miR-203a-3p expression levels were increased in pancreatic cancer Bxpc-3,Aspc-1 and Panc-1 cell lines,consistent with the results of TCGA database analysis;the expression level of miR-451 a was inconsistent with the analysis result of TCGA database(P<0.05).3 The results of validation of transfection validity showed that the expression level of miR-203a-3p was significantly increased after transfection of miR-203a-3p mimic compared with NC mimic group,the expression level of miR-203a-3p was significantly decreased after transfection of miR-203a-3p inhibitor compared with NC inhibitor group(P<0.05).4 CCK-8 experiments showed that compared with the transfected NC mimic group,the transfected miR-203a-3p mimic group promoted the proliferation of pancreatic cancer cell lines Bxpc-3,Panc-1(P<0.05).5 Transwell experiments show that compared with the transfected NC mimic group,overexpression of miR-203a-3p promotes the ability of pancreatic cancer cell lines Bxpc-3,Panc-1 to migrate and invade;blocking endogenous miR-203a-3p function,it inhibited the ability of pancreatic cancer cell lines to migrate and invade(P<0.05).6 The results of cell colony formation experiments showed that after transfection of miR-203a-3p mimic into pancreatic cancer cell lines,compared with the control of transfection,miR-203a-3p group enhanced cell colony forming ability;blocked endogenous miR-203a-3p expression,compared with the NC inhibitor group,the cell colony forming ability of the miR-203a-3p inhibitor group was reduced(P<0.05).7 Multi-site predictions show that miR-203a-3p directly targets the 3’UTR region of PPM1 A mRNA,gene interaction network analysis shows that PPM1 A interacts with multiple genes.Conclusions 1 miR-203a-3p is highly expressed in pancreatic cancer tissue samples of the TCGA database and has a wide range of species conservation,which is related to the clinical stage of patients.The high expression of miR-203a-3p may be related to the poor prognosis and survival of patients.2 miR-203a-3p can promote the proliferation,migration,invasion and colony formation of pancreatic cancer cell lines Bxpc-3 and Panc-1.3 Bioinformatics website suggests that PPM1 A is the target gene of miR-203a-3p and PPM1 A has strong interaction with SMAD3,SMAD2 and SMAD4.Figure 28;Table 4;Reference 130.