BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found t...BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.展开更多
【目的】探明COL1A1(Collagen type I alpha 1 chain,I型胶原蛋白α1链)和COL1A2(Collagen type I alpha 2 chain,I型胶原蛋白α2链)基因在梅花鹿不同组织中的表达谱,解析其对梅花鹿组织发育的影响,为影响梅花鹿重要经济性状的候选基因...【目的】探明COL1A1(Collagen type I alpha 1 chain,I型胶原蛋白α1链)和COL1A2(Collagen type I alpha 2 chain,I型胶原蛋白α2链)基因在梅花鹿不同组织中的表达谱,解析其对梅花鹿组织发育的影响,为影响梅花鹿重要经济性状的候选基因筛选提供依据。【方法】采用RT-qPCR方法检测COL1A1和COL1A2基因在雄性梅花鹿心脏、肝脏和脾脏等16个组织器官中的表达水平;结合NetPhos 3.1、Motif Search和ProtParam等系列软件预测分析COL1A1和COL1A2基因的生物信息及其在梅花鹿不同组织中的表达谱,并在此基础上构建COL1A1和COL1A2氨基酸序列的系统发育进化树。【结果】COL1A1和COL1A2基因CDS区分别编码1463和1364个氨基酸,理论PI分别为5.60和9.19,COL1A1和COL1A2均是一种具有信号肽和磷酸化位点的亲水性稳定蛋白质;二者蛋白二级及三级结构均以无规则卷曲构成;与其他动物相比,鹿COL1A1和COL1A2基因均与反刍动物山羊、绵羊和牛的同源性最高,其中,鹿COL1A1基因与山羊、牛、绵羊的同源性分别为99.5%、99.5%和99.2%,鹿COL1A2基因与牛、绵羊、山羊的同源性分别为99.1%、99.0%和98.9%,亲缘关系最近。RT-qPCR结果显示,COL1A1和COL1A2基因在梅花鹿不同组织中均有表达,其中COL1A1基因在心脏、背最长肌和腿肌中的表达较高,显著高于其他组织,COL1A2基因在心脏、肝脏、肾脏和瓣胃中的表达较高,均显著高于其他组织;此外,COL1A1在肌肉组织中的表达较高,COL1A2较低;二者在其余组织中的表达具有一高一低,相互协同的作用趋势。【结论】COL1A1和COL1A2可能通过相互协同共同维持组织结构及组织发育,相关结果为后续深入研究COL1A1和COL1A2影响梅花鹿生长发育奠定基础。展开更多
文摘BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.
文摘【目的】探明COL1A1(Collagen type I alpha 1 chain,I型胶原蛋白α1链)和COL1A2(Collagen type I alpha 2 chain,I型胶原蛋白α2链)基因在梅花鹿不同组织中的表达谱,解析其对梅花鹿组织发育的影响,为影响梅花鹿重要经济性状的候选基因筛选提供依据。【方法】采用RT-qPCR方法检测COL1A1和COL1A2基因在雄性梅花鹿心脏、肝脏和脾脏等16个组织器官中的表达水平;结合NetPhos 3.1、Motif Search和ProtParam等系列软件预测分析COL1A1和COL1A2基因的生物信息及其在梅花鹿不同组织中的表达谱,并在此基础上构建COL1A1和COL1A2氨基酸序列的系统发育进化树。【结果】COL1A1和COL1A2基因CDS区分别编码1463和1364个氨基酸,理论PI分别为5.60和9.19,COL1A1和COL1A2均是一种具有信号肽和磷酸化位点的亲水性稳定蛋白质;二者蛋白二级及三级结构均以无规则卷曲构成;与其他动物相比,鹿COL1A1和COL1A2基因均与反刍动物山羊、绵羊和牛的同源性最高,其中,鹿COL1A1基因与山羊、牛、绵羊的同源性分别为99.5%、99.5%和99.2%,鹿COL1A2基因与牛、绵羊、山羊的同源性分别为99.1%、99.0%和98.9%,亲缘关系最近。RT-qPCR结果显示,COL1A1和COL1A2基因在梅花鹿不同组织中均有表达,其中COL1A1基因在心脏、背最长肌和腿肌中的表达较高,显著高于其他组织,COL1A2基因在心脏、肝脏、肾脏和瓣胃中的表达较高,均显著高于其他组织;此外,COL1A1在肌肉组织中的表达较高,COL1A2较低;二者在其余组织中的表达具有一高一低,相互协同的作用趋势。【结论】COL1A1和COL1A2可能通过相互协同共同维持组织结构及组织发育,相关结果为后续深入研究COL1A1和COL1A2影响梅花鹿生长发育奠定基础。