MicroRNA-140-5p Suppresses Cell Proliferation And Invasion Ability In Esophageal Squamous Cell Carcinoma By Targeting Glut1

Esophageal carcinoma is the eighth most common carcinoma in the world.It has a high lethality rate,with more than 400,000 deaths per year.Esophageal squamous cell carcinoma(ESCC)is the most common type of esophageal carcinoma globally and in economically developing countries.Approximately half of the world’s 0.5millin new ESCC cases annually occur in China,where the disease is a major public health problem.With insidious early symptoms,patients with ESCC often have advanced stage disease at initial diagnosis.Micro RNAs(miRNAs)are small noncoding RNA sequences of 19–25nucleotides that play a role in the post-transcriptional control of gene expression.Recently,it has been reported that miRNAs are involved in the control of cell proliferation,apoptosis,cell cycle distribution,cellular migration and angiogenesis.Accumulating evidence shows that miRNAs are often abnormally expressed in diverse cancers,including leukemias and lymphomas,colorectal cancer,liver cancer,head and neck cancer and breast cancers.Glucose transporter 1(GLUT1),the uniporter protein encoded by the SLC2A1 gene,is a key rate-limiting factor in the transport of glucose in cancer cells,and frequently expressed in a significant proportion of human cancers.Malignant cells are known to reprogram their metabolism to boost the rapid growth,proliferation and long-lasting maintenance.In light of the promoting role of GLUT1 in tumor metabolism and development,targeting GLUT1 for therapeutics and prevention might be conducive.The function of miR-140-5p in ESCC is currently unknown.In this study,we explored the expression of miR-140-5p in ESCC tissues and cells,and analyzed the effect of its expression on ESCC cell proliferation and invasion.Furthermore,a double luciferase reporting experiment was used to confirm that glucose transporter 1(Glut1)is a direct target of miR-140-5p,and it has a negative correlation with miR-140-5p expression in ESCC tissues.These data suggest that miR-140-5p is closely related to the occurrence and development of ESCC,and targeting miR-140-5p/Glut1 is expected to become a new treatment strategy for ESCC patients.Methods1.Expression and clinical significance of miR-140-5p in esophageal squamous cell carcinoma(1)GEO data analysis of miR-140-5p expression in ESCC tissues and normal tissues.(2)Real-time quantitative PCR(q PCR)analysis of miR-140-5p expression in ESCC tissues and cells.2.miR-140-5p affects the proliferation and migration of esophageal squamous cell carcinoma cells(1)CCK-8 method was used to detect the proliferation of esophageal squamous cell carcinoma cells after transfection.(2)Transwell method was used to detect the invasive ability of esophageal squamous cell carcinoma cells after transfection.3.miR-140-5p regulates the biological behavior of esophageal squamous carcinoma cells through GUT1(1)Dual-Luciferase Reporter assay was used to verify the interaction of miR-140-5p with Glut1.(2)Western blot was utilized to detect the expression of Glut1 protein in Eca109 and KYSE70 cells after transfection4.Statistical analysisGraph Pad Prism 6.0 software was used for statistical analysis.The measurement data is expressed as the comparison between the two groups using the t test,the comparison between the three groups and more samples using the single factor analysis of variance,the correlation analysis using the Pearson correlation analysis method,the test level α=0.05Results1.miR-140-5p is underexpressed in esophageal squamous cell carcinoma(1)GEO datasets assay revealed that the expression of miR-140-5p in ESCC tissues(0.205±0.201)was significantly lower than that in normal tissues(0.941±0.364),and the difference was statistical significance(P<0.05).(2)The result of q PCR demonstrated that the expression of miR-140-5p in ESCC tissues and cells were markedly lower than that in normal tissues and normal esophageal epithelial cell Het-1A,and statistical difference was found(P<0.01).2.miR-140-5p affects esophageal squamous cell carcinoma cell proliferation and migration(1)miR-140-5p mimic significantly upregulated the expression of miR-140-5p in Eca109 and KYSE70 cells,and its upregulation suppressed cell proliferation in Eca109 and KYSE70 cells.(2)miR-140-5p mimic significantly upregulated the expression of miR-140-5p in Eca109 and KYSE70 cells,and its upregulation suppressed cell invasion in Eca109 and KYSE70 cells.3.miR-140-5p regulates the biological behavior of esophageal squamous carcinoma cells through GUT1(1)Western blot results showed that compared with the negative control group,the expression of Glut1 protein in the miR-140-5p analog group Eca109 and KYSE70 cells was significantly down-regulated,and the difference was statistical significance(P<0.05).(2)Dual-Luciferase Reporter assay indicated that Glut1 was a direct target of miR-140-5p in ESCC cells,and its expression was upregulated in ESCC tissues.Glut1 expression displayed obvious negative correlation with miR-140-5p expression in ESCC tissues.MiR-140-5p mimic dramatically inhibited the expression of Glut1 in Eca109 and KYSE70 cells.ConclusionMiR-140-5p plays an important role in the occurrence and development of ESCC.It can inhibit the proliferation and invasion of esophageal squamous cell carcinoma by targeting Glut1.Targeting miR-140-5p/Glut1 may be one of the new treatment strategies for ESCC patients.