| Objective:In the TCGA database,we obtained differentially expressed IncRNA through bioinformatics analysis,then we screen out the lncRNA TM4SF1-AS1 we want to study.This study aimed to explore the expression and biological significance of TM4SF1-AS1 in colorectal cancer.Methods:Download TCGA database colorectal cancer HTSeq-FPKM transcript data,summarize the data through perl language,use the R language edgeR package to analyze the difference between 647 lncRNA transcripts of colorectal cancer tumor tissues and 51 IncRNA transcripts of tissues adjacent to cancer,get differentially expressed lncRNAs,the differentially expressed lncRNAs were screened for TM4SF1-AS1 by Wilcoxon rank sum test and the correlation with clinical stage and the specificity and sensitivity analysis of colorectal cancer tissue relative to tissues adjacent to cancer,the expression of TM4SF1-AS1 in all colorectal cancer patients was divided into high and low expression groups by the median value,Kaplan-Meier method and log-rank test was used for survival analysis;Collect the information of 99 colorectal cancer patients and their tumor tissue samples and matched paracancerous tissue samples,detection and analysis of the expression level of TM4SF1-AS1 in paired cancer tissues and adjacent tissues by qRT-PCR technology,then Kaplan-Meier survival analysis and log-rank test were used to analyze the correlation between TM4SF1-AS1 expression and the prognosis of colorectal cancer patients.RT-PCR detects the expression of lncRNA-TM4SF1-AS1 in colorectal cancer cell lines(HCT116,HT29,SW480,SW620)and human normal colon epithelial cell line(FHC);Select colorectal cancer cell lines with higher expression of TM4SF1-AS1(HT29,SW620),transfect TM4SF1-AS1-siRNA into them to knock down the expression of TM4SF1-AS1,and verify transfection efficiency;The effects of down-regulated TM4SF1-AS1 expression on the proliferation of HT29 and SW620 cells were detected by colony formation assay.The effects of down-regulated TM4SF1-AS 1 expression on the migration and invasion of HT29 and SW620 cells were detected by scratch test and transwell invasion test.Results:Differential expression analysis of 647 colorectal cancer tumor tissue samples and 51 tissues adjacent to cancer samples lncRNA through R language edgeR package showed 1229 differentially expressed lncRNA,278 with high expression and 951 with low expression;647 tumor tissue samples and 51 tissues adjacent to cancer samples of TM4SF1-AS1 in TCGA were tested by Wilcoxon rank sum test in R language,theresults showed that the expression of TM4SF1-AS1 in tumor tissues was higher than that in tissues adjacent to cancer.The expression of TM4SF1-AS1 in tumor tissue samples of patients with ? and ? stage was higher than that of patients with I and? stage;Detect the sensitivity and specificity of TM4SF1-AS1 expression in tumor tissue compared with that in tissues adjacent to cancer,and the results showed that the specificity and sensitivity of TM4SF1-AS1 were 0.508 and 0.668 respectively,and the AUC value was 0.71;The OS and DFS of colorectal cancer patients with high expression of TM4SF1-AS1 were worse than those with low expression of TM4SF1-AS1.Analysis of 99 colorectal cancer clinical tissue samples found that compared with tissues adjacent to cancer,TM4SF1-AS1 is highly expressed in colorectal cancer tumor tissues and predicts shorter overall survival time and disease-free survival time;Univariate and multivariate Cox regression models indicate that TM4SF1-AS1 expression is an independent prognostic factor for colorectal cancer.RT-PCR detection showed that the relative expression of TM4SF1-AS1 in human colorectal cancer cell lines(HCT116,SW620,SW480,HT29)was higher than that in human normal colon epithelial cell line(FHC);TM4SF1-AS1 siRNA down-regulated the expression of TM4SF1-AS1 in the HT29 and SW620 cell lines;Colony formation assay proved that the down-regulation of TM4SF1-AS1 expression inhibited the proliferation of colorectal cancer cells HT29,SW620.Cell scratch test and Transwell invasion test show the down-regulation of TM4SF1-AS1 expression inhibits the migration and invasion of colorectal cancer cells HT29,SW620.Conclusions:LncRNA TM4SF1-AS1 plays a role in promoting tumors in colorectal cancer and is a potential prognostic biomarker in colorectal cancer. |
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