BACKGROUND Colorectal cancer(CRC)is the third most common cancer and the second most common cause of cancer-related death worldwide.The 5-year survival rate of patients with early-stage CRC could reach 90%,but it is v...BACKGROUND Colorectal cancer(CRC)is the third most common cancer and the second most common cause of cancer-related death worldwide.The 5-year survival rate of patients with early-stage CRC could reach 90%,but it is very low in patients with advanced-stage CRC.Recent studies have shown that circular RNAs play important roles in regulating the migration and invasion of CRC cells.AIM To elucidate the role of circRNA_0084927(circ_0084927)in the migration and invasion of CRC cells and its underlying mechanism.METHODS Clinical tissue samples and cells were collected,and the expression of circ_0084927 was detected by quantitative polymerase chain reaction(qPCR).The diagnostic performance of circ_0084927 was assessed by receiver operating characteristic curve analysis.The role of circ_0084927 in CRC cell proliferation,migration,and invasion was determined using cell counting kit-8 assay,wound healing assay,and transwell assay,respectively.The regulatory relationship among circ_0084927,miRNA-20b-3p(miR-20b-3p),and glutathione S-transferase mu 5(GSTM5)was identified using databases,luciferase reporter assay,qPCR,and Western blot analysis.AKT-mTOR signaling was also verified after circ_0084927 knockdown or miR-20b-3p mimic treatment.RESULTS The expression of circ_0084927 was significantly increased in CRC tissues and cells,and it was higher in advanced-stage CRC compared with early-stage CRC.The area under the curve(AUC)of circ_0084927 was 0.806[95%confidence interval(CI):0.683-0.896].In addition,the AUC was 0.874(95%CI:0.738-0.956)in patients with advanced-stage CRC and 0.713(95%CI:0.555-0.840)in those with early-stage CRC.Knockdown of circ_0084927 inhibited the migration and invasion of HCT116 cells.Moreover,circ_0084927 was found to act as a sponge of miR-20b-3p.MiR-20b-3p activation reduced the circ_0084927 level,whereas miR-20b-3p inhibition increased the circ_0084927 level.But the effect was not found after circ_0084927 mutation.In addition,miR-20b-3p expression in CRC patients was also reduced and negatively correlated with circ_0084927 expression.The function of circ_0084927 in HCT116 cells with circ_0084927 knockdown was rescued by miR-20b-3p.Moreover,GSTM5 expression was significantly decreased after overexpressing miR-20b-3p or inhibiting circ_0084927,but its expression was rescued when circ_0084927 and miR-20b-3p were both inhibited.Finally,AKTmTOR signaling was markedly regulated by circ_0084927 and miR-20b-3p.CONCLUSION The expression of circ_0084927 is significantly increased in CRC and higher in advanced-stage CRC than in early-stage CRC.Moreover,circ_0084927 potentially regulates CRC cell migration and invasion via the miR-20b-3p/GSTM5/AKT/mTOR pathway.展开更多
[目的]探讨H9c2心肌细胞肥大的潜在机制。[方法]苯肾上腺素处理后,检测H9c2细胞肥大情况。RNA-Seq检测H9c2细胞肥大后的差异miRNA。过表达差异miRNA,鉴定调控H9c2细胞肥大的miRNA。通过miRDB在线分析miRNA的潜在底物。[结果]苯肾上腺素...[目的]探讨H9c2心肌细胞肥大的潜在机制。[方法]苯肾上腺素处理后,检测H9c2细胞肥大情况。RNA-Seq检测H9c2细胞肥大后的差异miRNA。过表达差异miRNA,鉴定调控H9c2细胞肥大的miRNA。通过miRDB在线分析miRNA的潜在底物。[结果]苯肾上腺素处理H9c2细胞后,细胞表面积明显增大(1 467±123 vs 3 764±312,P<0.05),并且通过RNA-Seq发现多种miRNA的表达水平下降。当过表达rno-miR-301b-3p时,H9c2细胞的表面积减少(3 427±252 vs 1 788±120,P<0.05)。敲低rno-miR-301b-3p时,H9c2细胞的表面积增加(1 542±243 vs 2 890±111,P<0.05)。过表达rno-miR-301b-3p后,发现FBXO48、SLC9A4、CHST1、UBL3基因的mRNA表达水平显著下降。敲低SLC9A4后,H9c2细胞的表面积减少(3 651±135 vs 1 777±124,P<0.05)。敲低rno-miR-301b-3p后,SLC26A4的蛋白表达水平上升。过表达rno-miR-301b-3p后,SLC9A4的蛋白表达水平下降。过表达SLC9A4后,H9c2细胞的表面积增加(1 456±111 vs 3 324±321,P<0.05)。此外,苯肾上腺素处理H9c2细胞后,SLC26A4的蛋白表达水平上升。[结论] rno-miR-301b-3p通过靶向SLC26A4 mRNA的3′端非编码区,降解了SLC26A4的mRNA,抑制了SLC26A4的蛋白表达,最后抑制了H9c2细胞肥大。展开更多
文摘BACKGROUND Colorectal cancer(CRC)is the third most common cancer and the second most common cause of cancer-related death worldwide.The 5-year survival rate of patients with early-stage CRC could reach 90%,but it is very low in patients with advanced-stage CRC.Recent studies have shown that circular RNAs play important roles in regulating the migration and invasion of CRC cells.AIM To elucidate the role of circRNA_0084927(circ_0084927)in the migration and invasion of CRC cells and its underlying mechanism.METHODS Clinical tissue samples and cells were collected,and the expression of circ_0084927 was detected by quantitative polymerase chain reaction(qPCR).The diagnostic performance of circ_0084927 was assessed by receiver operating characteristic curve analysis.The role of circ_0084927 in CRC cell proliferation,migration,and invasion was determined using cell counting kit-8 assay,wound healing assay,and transwell assay,respectively.The regulatory relationship among circ_0084927,miRNA-20b-3p(miR-20b-3p),and glutathione S-transferase mu 5(GSTM5)was identified using databases,luciferase reporter assay,qPCR,and Western blot analysis.AKT-mTOR signaling was also verified after circ_0084927 knockdown or miR-20b-3p mimic treatment.RESULTS The expression of circ_0084927 was significantly increased in CRC tissues and cells,and it was higher in advanced-stage CRC compared with early-stage CRC.The area under the curve(AUC)of circ_0084927 was 0.806[95%confidence interval(CI):0.683-0.896].In addition,the AUC was 0.874(95%CI:0.738-0.956)in patients with advanced-stage CRC and 0.713(95%CI:0.555-0.840)in those with early-stage CRC.Knockdown of circ_0084927 inhibited the migration and invasion of HCT116 cells.Moreover,circ_0084927 was found to act as a sponge of miR-20b-3p.MiR-20b-3p activation reduced the circ_0084927 level,whereas miR-20b-3p inhibition increased the circ_0084927 level.But the effect was not found after circ_0084927 mutation.In addition,miR-20b-3p expression in CRC patients was also reduced and negatively correlated with circ_0084927 expression.The function of circ_0084927 in HCT116 cells with circ_0084927 knockdown was rescued by miR-20b-3p.Moreover,GSTM5 expression was significantly decreased after overexpressing miR-20b-3p or inhibiting circ_0084927,but its expression was rescued when circ_0084927 and miR-20b-3p were both inhibited.Finally,AKTmTOR signaling was markedly regulated by circ_0084927 and miR-20b-3p.CONCLUSION The expression of circ_0084927 is significantly increased in CRC and higher in advanced-stage CRC than in early-stage CRC.Moreover,circ_0084927 potentially regulates CRC cell migration and invasion via the miR-20b-3p/GSTM5/AKT/mTOR pathway.
文摘[目的]探讨H9c2心肌细胞肥大的潜在机制。[方法]苯肾上腺素处理后,检测H9c2细胞肥大情况。RNA-Seq检测H9c2细胞肥大后的差异miRNA。过表达差异miRNA,鉴定调控H9c2细胞肥大的miRNA。通过miRDB在线分析miRNA的潜在底物。[结果]苯肾上腺素处理H9c2细胞后,细胞表面积明显增大(1 467±123 vs 3 764±312,P<0.05),并且通过RNA-Seq发现多种miRNA的表达水平下降。当过表达rno-miR-301b-3p时,H9c2细胞的表面积减少(3 427±252 vs 1 788±120,P<0.05)。敲低rno-miR-301b-3p时,H9c2细胞的表面积增加(1 542±243 vs 2 890±111,P<0.05)。过表达rno-miR-301b-3p后,发现FBXO48、SLC9A4、CHST1、UBL3基因的mRNA表达水平显著下降。敲低SLC9A4后,H9c2细胞的表面积减少(3 651±135 vs 1 777±124,P<0.05)。敲低rno-miR-301b-3p后,SLC26A4的蛋白表达水平上升。过表达rno-miR-301b-3p后,SLC9A4的蛋白表达水平下降。过表达SLC9A4后,H9c2细胞的表面积增加(1 456±111 vs 3 324±321,P<0.05)。此外,苯肾上腺素处理H9c2细胞后,SLC26A4的蛋白表达水平上升。[结论] rno-miR-301b-3p通过靶向SLC26A4 mRNA的3′端非编码区,降解了SLC26A4的mRNA,抑制了SLC26A4的蛋白表达,最后抑制了H9c2细胞肥大。
