MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to...MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to provide a theoretical basis for treatment.Bioinformatics was used to analyze pituitary adenoma-related genes and screen new targets related to RECK and miRNA.As well,the relationship between miR-200b-3p and RECK protein was verified using a double-luciferase reporter gene assay.The expression of miR-200b-3p in clinical samples was analyzed by in situ hybridization.Transfection of the miR-200b-3p inhibitor and small interfering-RECK(si-RECK)was verified by qPCR.GH3 cell viability and proliferation were detected using CCK8 and EdU assays.Apoptosis was detected by flow cytometry and western blotting.Wound healing and Transwell assays were used to detect cell migration and invasion.The effects of miR-200b-3p and RECK on GH3 cells were verified using salvage experiments.miR-200b-3p was highly expressed in pituitary tumor tissue.Inhibitors of miR-200b-3p inhibited cell proliferation promoted cell apoptosis,inhibited invasion and migration,and inhibited the expression of matrix metalloproteinases.Interestingly,miR-200b-3p negatively regulated RECK.The expression of RECK in pituitary adenoma tissues was lower than that in neighboring tissues.Si-RECK rescued the function of miR-200b-3p inhibitors in the above cellular behaviors,and miR-200b-3p accelerated the development of pituitary adenoma by negatively regulating RECK expression.In summary,this study investigated the molecular mechanism by which miR-200b-3p regulates the progression of pituitary adenoma through the negative regulation of RECK.The findings provide a new target for the treatment of pituitary adenoma.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
基金supported by Correlation between RECK and GH-type pituitary adenomas(No.21JR11RE027).
文摘MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to provide a theoretical basis for treatment.Bioinformatics was used to analyze pituitary adenoma-related genes and screen new targets related to RECK and miRNA.As well,the relationship between miR-200b-3p and RECK protein was verified using a double-luciferase reporter gene assay.The expression of miR-200b-3p in clinical samples was analyzed by in situ hybridization.Transfection of the miR-200b-3p inhibitor and small interfering-RECK(si-RECK)was verified by qPCR.GH3 cell viability and proliferation were detected using CCK8 and EdU assays.Apoptosis was detected by flow cytometry and western blotting.Wound healing and Transwell assays were used to detect cell migration and invasion.The effects of miR-200b-3p and RECK on GH3 cells were verified using salvage experiments.miR-200b-3p was highly expressed in pituitary tumor tissue.Inhibitors of miR-200b-3p inhibited cell proliferation promoted cell apoptosis,inhibited invasion and migration,and inhibited the expression of matrix metalloproteinases.Interestingly,miR-200b-3p negatively regulated RECK.The expression of RECK in pituitary adenoma tissues was lower than that in neighboring tissues.Si-RECK rescued the function of miR-200b-3p inhibitors in the above cellular behaviors,and miR-200b-3p accelerated the development of pituitary adenoma by negatively regulating RECK expression.In summary,this study investigated the molecular mechanism by which miR-200b-3p regulates the progression of pituitary adenoma through the negative regulation of RECK.The findings provide a new target for the treatment of pituitary adenoma.
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。