rno-miR-301b-3p靶向SLC26A4调控H9c2细胞肥大

rno-miR-301b-3p targeting SLC26A4 in regulating myocardial hypertrophy of H9c2 cells

[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。

[Objective]To explore the underlying mechanism of myocardial hypertrophy in H9 c2 cells.[Method] After phenylephrine treatment,the myocardial hypertrophy of H9 c2 cells was detected.The differential miRNAs of H9 c2 cells after myocardial hypertrophy were detected by RNA-Seq.Through overexpression of differential miRNAs,miRNAs that regulate myocardial hypertrophy of H9 c2 cells were identified.Online analysis of potential substrates of miRNA through miRDB.[Result] H9 c2 cells treated with phenylephrine significantly increased cell surface area(1 467±123 vs 3 764±312,P<0.05),and the expression levels of various miRNAs were decreased by RNA-seq.When rno-miR-301 b-3 p was overexpressed,the myocardial surface area of H9 c2 cells decreased(3 427±252 vs 1 788±120,P<0.05).When rno-miR-301 b-3 p was knocked down,the myocardial surface area of H9 c2 cells increased(1 542±243 vs 2 890±111,P<0.05).After the overexpression of rno-miR-301 b-3 p,the mRNA expression levels of FBXO48,SLC9 A4,CHST1 and UBL3 genes were significantly decreased.After SLC9 A4 knockdown,the myocardial surface area of H9 c2 cells decreased(3 651±135 vs 1 777±124,P<0.05).After knockdown of rno-miR-301 b-3 p,the protein expression level of SLC26 A4 increased.After the overexpression of rno-miR-301 b-3 p,the protein expression level of SLC9 A4 decreased.After SLC9 A4 overexpression,the myocardial surface area of H9 c2 cells increased(1 456±111 vs 3 324±321,P<0.05).Furthermore,the protein expression level of SLC26 A4 was increased after H9 c2 cells were treated with phenylephrine.[Conclusion]rno-miR-301 b-3 p degrades the SLC26 A4 mRNA by targeting the 3′non-coding region of SLC26 A4 mRNA,inhibits the protein expression of SLC26 A4,and finally inhibits myocardial hypertrophy in H9 c2 cells.