Study On The Expression And Significance Of Long Non-coding RNA GAS6-AS1 In Colorectal Cancer

Objective:The purpose of this study is to find out the lncRNA which is differentially expressed in colorectal cancer(CRC)and has clinical significance,and to determine the effect of long-chain non coding RNA GAS6-AS1 on the proliferation,migration and invasion of colorectal cancer cells through further cell experiments in vitro.Method:1.In this study,we first collected and used the mRNA-seq data and corresponding follow-up information of colorectal cancer samples(including 647 colorectal cancer tissues and 51 adjacent normal tissues)retrieved and downloaded from TCGA database.Then,we use the edger package of R language to analyze the differentially expressed lncRNAs,and find out the significantly differentially expressed lncRNAs.Then we select the target IncRNA GAS6-AS1 by searching the literature related to these differentially expressed lncRNAs,and use R language to analyze the correlation between the target lncRNAs and the prognosis of colorectal cancer.2.The specimens of 30 patients with colorectal cancer were collected,and real-time quantitative PCR(RT-PCR)was used to detect the differential expression of lncRNA GAS6-AS1 in tumor tissues and normal control tissues.Similarly,the expression of lncRNA GAS6-AS1 in normal colorectal epithelial cell lines(FHC)and colorectal cancer cell lines(HCT116,SW480,SW620,HT29)was detected by real-time quantitative PCR.The synthesized siRNA was transfected into the relatively high expression colorectal cancer cells(HCT116).The cell lines were divided into negative control group(si-NC)and lncRNA GAS6-AS1 inhibition group(GAS6-AS1si1,GAS6-AS1si2,GAS6-AS1si3).RT-PCR was used to verify the transfection effect.After the cells were transfected successfully,the proliferation ability of cells was detected by clone plate test,the migration and invasion ability of cells were detected by cell scratch test and Transwell invasion testResult:(1)There were 1229 lncRNAs differentially expressed in colorectal cancer samples downloaded from TCGA database,of which 278 were up-regulated and 951 were down regulated,and the expression of GAS6-AS1 in colorectal cancer tissues was significantly higher than that in normal intestinal epithelial tissues(P<0.001);(2)The overall survival(OS)and disease free survival(DFS)of patients with high expression of GAS6-AS1 were significantly shorter than those of patients with low expression;(3)lncRNA GAS6-AS1 as a diagnostic indicator of colorectal cancer has high specificity and sensitivity,and has potential diagnostic value;(4)The tissue samples of 30 patients with colorectal cancer were detected by RT-PCR,and it was found that the expression of lncRNA GAS6-AS1 in cancer tissues was higher than that in adjacent tissues;(5)The expression of lncRNA GAS6-AS1 in four colorectal cancer cell lines was significantly higher than that in normal colon epithelial cells;(6)The results of clone plate experiment showed that the proliferation ability of tumor cells decreased significantly after down regulating the expression of lncRNA GAS6-AS1,and the invasion and migration ability of tumor cells decreased significantly after down regulating the expression of lncRNA GAS6-AS1 by scratch experiment and Transwell experiment;Conclusion:LncRNA GAS6-AS1 has a higher expression in colorectal cancer than that in normal tissues and cell;The higher expression of lncRNA GAS6-AS1is related to the poor prognosis of patients;LncRNA GAS6-AS1 has the ability to promote cell migration and invasion,and has the role of promoting oncogenes;...