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
为利用斑马鱼动物模型进一步研究bHLH转录因子家庭重要成员hand2基因在心脏发育中的功能,采用生物信息学结合RT-PCR的方法获得了斑马鱼hand2基因.将所得片段插入原核表达载体pGEX-4T-1中,并通过IPTG诱导表达出GST-Hand2融合蛋白,经GST...为利用斑马鱼动物模型进一步研究bHLH转录因子家庭重要成员hand2基因在心脏发育中的功能,采用生物信息学结合RT-PCR的方法获得了斑马鱼hand2基因.将所得片段插入原核表达载体pGEX-4T-1中,并通过IPTG诱导表达出GST-Hand2融合蛋白,经GST亲和层析法纯化后,免疫新西兰大白兔制备了多克隆抗体,利用western b lotting和免疫组化的方法对抗体进行分析.分析表明:斑马鱼hand2基因成熟肽编码区含有627 bp,编码208个氨基酸,与人Hand2蛋白的同源性达到83%;IPTG诱导表达后,表达的融合蛋白占菌体总蛋白的71%,经GST纯化获得SDS-PAGE电泳下单一条带.融合蛋白相对分子质量约为49 000.分析表明,制备的抗体具有很好的特异性.展开更多
目的通过Meta分析评估血液中lncRNA HAND2反义RNA1(HAND2-AS1)的表达水平对多种肿瘤的临床诊断价值。方法由两名评价员利用计算机检索PubMed、Embase、SinoMed和万方等数据库,检索时限为1980年1月1日至2020年9月30日,收集国内外公开发...目的通过Meta分析评估血液中lncRNA HAND2反义RNA1(HAND2-AS1)的表达水平对多种肿瘤的临床诊断价值。方法由两名评价员利用计算机检索PubMed、Embase、SinoMed和万方等数据库,检索时限为1980年1月1日至2020年9月30日,收集国内外公开发表有关HAND2-AS1表达水平与肿瘤癌症相关的所有文献,采用STATA 14.0软件对数据进行Meta分析。结果严格按照纳入和排除标准,筛选出6篇文献进行Meta分析,包括317名肿瘤患者和227名健康对照者。连续型变量所合并的效应量为标准化均数差(Standardized mean difference,SMD),具体为[SMD=-1.61,95%CI(-1.80,-1.41),P<0.00001]。对于HAND2-AS1的诊断价值,汇总敏感度、特异度分别为0.87和0.82,诊断比值比(Diagnostic Odds Ratio,DOR)为32,综合受试者工作特征曲线的曲线下面积(Aarea under curve,AUC)为0.91。结论血液中HAND2-AS1的检测在多种肿瘤的诊断中具有较高的敏感度和特异度,对区分患者和健康个体具有较高的临床诊断价值。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.
文摘为利用斑马鱼动物模型进一步研究bHLH转录因子家庭重要成员hand2基因在心脏发育中的功能,采用生物信息学结合RT-PCR的方法获得了斑马鱼hand2基因.将所得片段插入原核表达载体pGEX-4T-1中,并通过IPTG诱导表达出GST-Hand2融合蛋白,经GST亲和层析法纯化后,免疫新西兰大白兔制备了多克隆抗体,利用western b lotting和免疫组化的方法对抗体进行分析.分析表明:斑马鱼hand2基因成熟肽编码区含有627 bp,编码208个氨基酸,与人Hand2蛋白的同源性达到83%;IPTG诱导表达后,表达的融合蛋白占菌体总蛋白的71%,经GST纯化获得SDS-PAGE电泳下单一条带.融合蛋白相对分子质量约为49 000.分析表明,制备的抗体具有很好的特异性.
文摘目的通过Meta分析评估血液中lncRNA HAND2反义RNA1(HAND2-AS1)的表达水平对多种肿瘤的临床诊断价值。方法由两名评价员利用计算机检索PubMed、Embase、SinoMed和万方等数据库,检索时限为1980年1月1日至2020年9月30日,收集国内外公开发表有关HAND2-AS1表达水平与肿瘤癌症相关的所有文献,采用STATA 14.0软件对数据进行Meta分析。结果严格按照纳入和排除标准,筛选出6篇文献进行Meta分析,包括317名肿瘤患者和227名健康对照者。连续型变量所合并的效应量为标准化均数差(Standardized mean difference,SMD),具体为[SMD=-1.61,95%CI(-1.80,-1.41),P<0.00001]。对于HAND2-AS1的诊断价值,汇总敏感度、特异度分别为0.87和0.82,诊断比值比(Diagnostic Odds Ratio,DOR)为32,综合受试者工作特征曲线的曲线下面积(Aarea under curve,AUC)为0.91。结论血液中HAND2-AS1的检测在多种肿瘤的诊断中具有较高的敏感度和特异度,对区分患者和健康个体具有较高的临床诊断价值。
