| Background:Colorectal cancer is one of the most common malignant tumors in the world,and its morbidity and mortality rank third among all malignant tumors.Despite the advancement and optimization of treatment regimens,the treatment effect of colorectal cancer has been significantly improved in recent years,colorectal cancer patients are still associated with a poor five-year survival rate.Therefore,the search for early tumor diagnostic markers and new therapeutic targets will be an important clinical strategy to improve the survival rate of colorectal cancer patients.Long non-coding RNAs(lnc RNAs)are a class of RNAs with transcripts longer than 200 nt which lack a complete functional open reading frame(ORF).Lnc RNAs are abnormally expressed in various tumors and can be involved in tumor genesis by regulating functional proteins,which is an important marker and target for tumor diagnosis and treatment.Based on the analysis of colorectal cancer-related datasets in the GEO database,a new Lnc RNA LAMTOR5-AS1 related to colorectal cancer was screened in the early stage of this study.And the potential expression correlation between LAMTOR5-AS1 and ROCK1/ROCK2 was preliminarily found through the prediction of bioinformatics website.Objective:1.To explore the expression and clinical significance of LAMTOR5-AS1 in colorectal cancer tissues.2.To explore the correlation and clinical significance of LAMTOR5-AS1,ROCK1,and ROCK2 in colorectal cancer tissues.3.To analyze the expression and potential biological regulation of LAMTOR5-AS1 in colorectal cancer cells.Methods:1.Based on the colorectal cancer dataset of GEO database and bioinformatics website,the expression of LAMTOR5-AS1 in colorectal cancer and the relationship between its expression and clinicopathological features were further analyzed.2.139 pairs of colorectal cancer tissues and matched normal mucosa tissues were collected.The expression of LAMTOR5-AS1 was detected by in situ hybridization experiment,and the expression of ROCK1 and ROCK2 was detected by immunohistochemical experiment.3.The relationship between LAMTOR5-AS1,ROCK1 and ROCK2 expression and clinicopathological characteristics of colorectal cancer patients was analyzed.At the same time,the relationship between the co-expression of LAMTOR5-AS1 and ROCK1 or ROCK2 and the clinicopathological characteristics of colorectal cancer patients was analyzed.4.The expression of endogenous LAMTOR5-AS1 in colorectal cancer cells was inhibited by si RNA technology,and the m RNA and protein expressions of ROCK1 and ROCK2 were detected by real-time fluorescent quantitative PCR and Western blot experiments.5.The effects of LAMTOR5-AS1 on the migration of colorectal cancer were investigated by scratch and transwell assay.Results:1.The analysis of the bioinformatics website showed that the expression of LAMTOR5-AS1 was positively correlated with the metastasis and stage of colorectal cancer patients(p<0.05).The disease-free survival of patients with low LAMTOR5-AS1 expression was significantly higher than that of patients with high LAMTOR5-AS1 expression(p<0.05).2.In situ hybridization and immunohistochemical experiments showed that LAMTOR5-AS1,ROCK1 and ROCK2 were highly expressed in colorectal cancer tissues compared with normal mucosal tissues(p<0.05).Moreover,the expression of LAMTOR5-AS1 was correlated with the expression of ROCK1 or ROCK2(p<0.05).3.Analysis of the relationship between the examined molecules and clinicopathological characteristics of colorectal cancer patients found that: the expression of LAMTOR5-AS1 was positively correlated with the T stage,N stage and tumor size of colorectal patients(p<0.05);the high expression of ROCK1 was associated with colorectal cancer.The N stage,nerve invasion and differentiation degree of patients were positively correlated(p<0.05);while the high expression of ROCK2 was positively correlated with T and N stages of colorectal patients(p<0.05).4.The co-positive expression of LAMTOR5-AS1 and ROCK1 was positively correlated with the age,differentiation degree and N stage of colorectal cancer patients;the co-positive expression of LAMTOR5-AS1 and ROCK2 was positively correlated with the tumor size and N stage of colorectal cancer patients.5.The results of real-time quantitative PCR and Western blot assays showed that the RNA and protein expression levels of ROCK1 and ROCK2 in HCT116 cells were significantly decreased after LAMTOR5-AS1 si RNA was transfected.6.Scratch and transwell assay showed that LAMTOR5-AS1 could promote the migration of colorectal cancer cells.Conclusion:1.LAMTOR5-AS1,ROCK1 and ROCK2 are highly expressed in colorectal cancer.When LAMTOR5-AS1 is co-positively expressed with ROCK1 or ROCK2,both of them are positively correlated with N stage in colorectal cancer patients,suggesting that LAMTOR5-AS1 can be used as a potential new diagnostic marker of colorectal cancer.2.LAMTOR5-AS1 may be involved in the development of colorectal cancer by regulating the expression of ROCK1 and ROCK2.15 pictures,20 tables,76 documents... |
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