Background:Chronic kidney disease(CKD)is associated with common pathophysiological processes,such as inflammation and fibrosis,in both the heart and the kidney.However,the underlying molecular mechanisms that drive th...Background:Chronic kidney disease(CKD)is associated with common pathophysiological processes,such as inflammation and fibrosis,in both the heart and the kidney.However,the underlying molecular mechanisms that drive these processes are not yet fully understood.Therefore,this study focused on the molecular mechanism of heart and kidney injury in CKD.Methods:We generated an microRNA(miR)-26a knockout(KO)mouse model to investigate the role of miR-26a in angiotensin(Ang)-II-induced cardiac and renal injury.We performed Ang-II modeling in wild type(WT)mice and miR-26a KO mice,with six mice in each group.In addition,Ang-II-treated AC16 cells and HK2 cells were used as in vitro models of cardiac and renal injury in the context of CKD.Histological staining,immunohistochemistry,quantitative real-time polymerase chain reaction(PCR),and Western blotting were applied to study the regulation of miR-26a on Ang-II-induced cardiac and renal injury.Immunofluorescence reporter assays were used to detect downstream genes of miR-26a,and immunoprecipitation was employed to identify the interacting protein of LIM and senescent cell antigen-like domain 1(LIMS1).We also used an adeno-associated virus(AAV)to supplement LIMS1 and explored the specific regulatory mechanism of miR-26a on Ang-II-induced cardiac and renal injury.Dunnett’s multiple comparison and t-test were used to analyze the data.Results:Compared with the control mice,miR-26a expression was significantly downregulated in both the kidney and the heart after Ang-II infusion.Our study identified LIMS1 as a novel target gene of miR-26a in both heart and kidney tissues.Downregulation of miR-26a activated the LIMS1/integrin-linked kinase(ILK)signaling pathway in the heart and kidney,which represents a common molecular mechanism underlying inflammation and fibrosis in heart and kidney tissues during CKD.Furthermore,knockout of miR-26a worsened inflammation and fibrosis in the heart and kidney by inhibiting the LIMS1/ILK signaling pathway;on the contrary,supplementation with exogenous miR-26a reversed all these changes.Conclusions:Our findings suggest that miR-26a could be a promising therapeutic target for the treatment of cardiorenal injury in CKD.This is attributed to its ability to regulate the LIMS1/ILK signaling pathway,which represents a common molecular mechanism in both heart and kidney tissues.展开更多
目的 探讨LIM和SH3蛋白1(LIM and SH3 protein 1, LASP1)、激肽释放酶相关肽酶11(Kallikrein related peptidase 11, KLK11)表达水平与结直肠癌(Colorectal cancer, CRC)临床病理的关系及预后影响因素。方法 选择2017年1月至2018年10月...目的 探讨LIM和SH3蛋白1(LIM and SH3 protein 1, LASP1)、激肽释放酶相关肽酶11(Kallikrein related peptidase 11, KLK11)表达水平与结直肠癌(Colorectal cancer, CRC)临床病理的关系及预后影响因素。方法 选择2017年1月至2018年10月于本院就诊并接受手术治疗的82例CRC患者作为研究对象,术中收集癌组织和癌旁组织。免疫组织化学法检测癌旁组织和癌组织中LASP1和KLK11的阳性表达情况,分析LASP1和KLK11阳性表达与CRC临床病理的相关性。ROC曲线分析LASP1和KLK11在CRC诊断中的价值,分析LASP1和KLK11在CRC癌组织中表达的相关性。对患者进行5年随访并记录患者生存情况,确定LASP1和KLK11阳性表达与CRC患者生存的关系,Cox分析确定影响CRC患者生存的风险因素。结果 相对于癌旁组织,LASP1和KLK11在CRC癌组织中的阳性表达更高(P<0.05)。LASP1和KLK11表达与TNM分期和淋巴结转移有关(P均<0.05)。相对于Ⅰ/Ⅱ期和未发生淋巴结转移的患者,Ⅲ/Ⅳ期和发生淋巴结转移的患者中LASP1和KLK11表达为阳性的占比更高(P均<0.05)。ROC曲线分析结果显示,LASP1和KLK11能够作为CRC的有效诊断指标,且二者联合诊断价值更高。相关性分析结果显示,LASP1和KLK11在CRC癌组织中的表达呈正相关(r=0.33,P=0.003)。LASP1阳性和KLK11阳性患者的生存率显著低于LASP1阴性和KLK11阴性患者(P<0.05)。Cox多因素分析显示,分化程度、TNM分期、淋巴结转移、LASP1阳性表达、KLK11阳性表达是影响CRC患者预后的独立因素(P均<0.05)。结论 LASP1和KLK11阳性表达与CRC患者临床病理指标存在相关性,LASP1和KLK11表达或可作为CRC的有效诊断指标且与患者预后相关。展开更多
目的探究微小RNA-625-5p(miR-625-5p)是否可靶向LIM和SH3结构域蛋白1(LIM and SH3 protein 1,LASP1)影响结直肠癌(colorectal cancer,CRC)细胞的增殖、迁移及凋亡。方法CRC细胞转染后分为空白对照组、阴性对照组、miR-625-5p类似物组、m...目的探究微小RNA-625-5p(miR-625-5p)是否可靶向LIM和SH3结构域蛋白1(LIM and SH3 protein 1,LASP1)影响结直肠癌(colorectal cancer,CRC)细胞的增殖、迁移及凋亡。方法CRC细胞转染后分为空白对照组、阴性对照组、miR-625-5p类似物组、miR-625-5p抑制剂组、miR-625-5p类似物+LASP1组。进行荧光定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)检测各组LASP1、miR-625-5p的mRNA表达,双荧光素酶报告基因检测基因靶向结合关系,细胞计数试剂盒(cell counting kit,CCK)-8检测细胞活性,细胞平板克隆检测细胞克隆能力,流式细胞术检测细胞凋亡,Transwell法检测细胞迁移及侵袭,EDU实验测定细胞增殖,蛋白免疫印迹(Western blot)法检测LASP1及侵袭转移相关蛋白神经型钙黏蛋白(neural cadherin,N-cadherin)和上皮型钙黏蛋白(epithelial cadherin,E-cadherin)的表达情况。结果qPCR检测证明细胞转染成功,双荧光素酶报告基因检测到miR-625-5p可靶向结合LASP1。与阴性对照组比较,miR-625-5p类似物组的细胞活性和细胞克隆、迁移、侵袭、增殖数量及LASP1、N-cadherin表达水平降低(P<0.05),细胞凋亡率及E-cadherin表达水平上升(P<0.05),miR-625-5p抑制组上述指标的结果完全相反。与miR-625-5p类似物组比较,miR-625-5p类似物+LASP1组的细胞活性和细胞克隆、迁移、侵袭、增殖数量及LASP1蛋白表达水平升高(P<0.05),细胞凋亡率降低(P<0.01)。结论miR-625-5p可靶向LASP1抑制结直肠癌细胞的增殖、迁移,并促进凋亡。展开更多
目的分析双硫死亡(disulfidptosis)相关基因人PDZ和LIM域蛋白1(PDZ and LIM domain protein 1,PDLIM1)m RNA在多种肿瘤中的表达及作用。方法通过仙桃学术网站分析PDLIM1 mRNA的表达情况。利用仙桃学术网站和Sangerbox 3.0数据分析平台探...目的分析双硫死亡(disulfidptosis)相关基因人PDZ和LIM域蛋白1(PDZ and LIM domain protein 1,PDLIM1)m RNA在多种肿瘤中的表达及作用。方法通过仙桃学术网站分析PDLIM1 mRNA的表达情况。利用仙桃学术网站和Sangerbox 3.0数据分析平台探究PDLIM1在33种肿瘤中的诊断和预后能力。利用TISIDB数据库分析PDLIM1与临床分级和分期的相关性。在Sangerbox 3.0数据分析平台和Kaplan-Meier Plotter数据库中分析PDLIM1与肿瘤免疫相关性。通过STRING数据库和Cytoscape构建蛋白质互作网络(protein-protein interaction networks,PPI)。利用Sangerbox 3.0数据分析平台进行富集分析。最后利用GSCA(Gene Set Cancer Analysis)网站分析获得PDLIM1 mRNA表达与药物的敏感性。结果PDLIM1 mRNA在33种肿瘤中表达量存在异质性。PDLIM1在胆管癌(CHOL)、多形性胶质母细胞瘤(GBM)、肾透明细胞癌(KIRC)、肺腺癌(LUAD)、卵巢癌(OV)、胰腺癌(PAAD)、黑色素瘤(SKCM)和睾丸生殖细胞肿瘤(TGCT)中具有良好的诊断能力。PDLIM1在胶质瘤(GBMLGG)、脑低级别胶质瘤(LGG)、混合肾癌(KIPAN)、多形性胶质细胞瘤(GBM)、间皮瘤(MESO)、葡萄膜黑色素瘤(UVM)和肾上腺皮质癌(ACC)中高表达预后差,而在肉瘤中低表达预后差。PDLIM1 mRNA表达与头颈鳞状细胞癌(HIVSC)、肾乳头状细胞癌(KIRP)、子宫内膜癌(UCEC)、子宫癌肉瘤(UCS)和葡萄膜黑色素瘤(UVM)的分级,以及与宫颈癌、头颈鳞状细胞癌、子宫内膜癌和脑低级别胶质瘤肿瘤的分期有关。PDLIM1与以前列腺癌为首的36种肿瘤的免疫浸润显著相关,且发现在PDLIM1 mRNA高表达的患者中经免疫治疗后的预后相对较好。PDLIM1在生物体内主要通过参与肌动蛋白细胞骨架、细胞黏附、肿瘤相关途径的调节来发挥作用,对以Isoliquiritigenin为首的多种药物敏感。结论PDLIM1与多种肿瘤的临床预后和免疫浸润等方面密切相关,有望成为一种肿瘤诊断和预后生物标志物或治疗靶点。展开更多
Background:Papillary thyroid cancer(PTC)is the most prevalent histological type of differentiated thyroid malignancy.Circular RNAs(circRNAs)have been implicated in the pathogenesis and progression of various cancers.c...Background:Papillary thyroid cancer(PTC)is the most prevalent histological type of differentiated thyroid malignancy.Circular RNAs(circRNAs)have been implicated in the pathogenesis and progression of various cancers.circTIAM1(hsa_circ_0061406)is a novel circRNA with aberrant expression in PTC.However,its functional roles in PTC progression remain to be investigated.Methods:The expression levels of circTIAM1 in the PTC and the matched para-cancerous tissues were detected by quantitative real-time reverse-transcription PCR(qRT-PCR).The subcellular localization of circTIAM1 was examined by fluorescence in-situ hybridization(FISH).Kaplan-Meier plot was used to analyze the association of clinicopathological features with circTIAM1 expression.Bioinformatics databases were utilized to predict the target miRNAs of circTIAM1 and the downstream target mRNAs.RNA pulldown,RIP assay,and dual-luciferase reporter assay were used to confirm the interactions.Functional experiments,such as CCK-8,EDU staining,and apoptosis assays,as well as in vivo xenograft model were employed to explore the impacts of circTIAM1,miR-338-3p,and LIM/SH3 protein 1(LASP1)on the malignant phenotype of the PTC cells.Results:CircTIAM1 was highly expressed in PTC cells.Moreover,circTIAM1 silencing suppressed the proliferation and invasion of PTC cells in vitro and impaired tumorigenesis in vivo.Furthermore,miR-338-3p was verified as a miRNA target of circTIAM1.LASP1 was also identified as a downstream target of miR-338-3p.The anti-tumorigenic effect of miR-338-3p overexpression and the pro-tumorigenic effect of LASP1 was further explored by functional assays,which demonstrated that circTIAM1 modulated the PTC progression through targeting miR-338-3p/LASP1 axis.Conclusion:The overexpression of circTIAM1 is associated with the malignant progression of PTC.A high level of circTIAM1 promotes the malignancy of PTC cells via the miR-338-3p/LASP1 axis.展开更多
基金supported by grants from the National Natural Science Foundation of China(Nos.82200749,82241047,82070735,82030024,81720108007 and 81270725)Natural Science Foundation of Jiangsu Province(No.BK20221282)National Key Research Programme of Ministry of Science and Technology(Nos.2018YFC130046,2018YFC1314000).
文摘Background:Chronic kidney disease(CKD)is associated with common pathophysiological processes,such as inflammation and fibrosis,in both the heart and the kidney.However,the underlying molecular mechanisms that drive these processes are not yet fully understood.Therefore,this study focused on the molecular mechanism of heart and kidney injury in CKD.Methods:We generated an microRNA(miR)-26a knockout(KO)mouse model to investigate the role of miR-26a in angiotensin(Ang)-II-induced cardiac and renal injury.We performed Ang-II modeling in wild type(WT)mice and miR-26a KO mice,with six mice in each group.In addition,Ang-II-treated AC16 cells and HK2 cells were used as in vitro models of cardiac and renal injury in the context of CKD.Histological staining,immunohistochemistry,quantitative real-time polymerase chain reaction(PCR),and Western blotting were applied to study the regulation of miR-26a on Ang-II-induced cardiac and renal injury.Immunofluorescence reporter assays were used to detect downstream genes of miR-26a,and immunoprecipitation was employed to identify the interacting protein of LIM and senescent cell antigen-like domain 1(LIMS1).We also used an adeno-associated virus(AAV)to supplement LIMS1 and explored the specific regulatory mechanism of miR-26a on Ang-II-induced cardiac and renal injury.Dunnett’s multiple comparison and t-test were used to analyze the data.Results:Compared with the control mice,miR-26a expression was significantly downregulated in both the kidney and the heart after Ang-II infusion.Our study identified LIMS1 as a novel target gene of miR-26a in both heart and kidney tissues.Downregulation of miR-26a activated the LIMS1/integrin-linked kinase(ILK)signaling pathway in the heart and kidney,which represents a common molecular mechanism underlying inflammation and fibrosis in heart and kidney tissues during CKD.Furthermore,knockout of miR-26a worsened inflammation and fibrosis in the heart and kidney by inhibiting the LIMS1/ILK signaling pathway;on the contrary,supplementation with exogenous miR-26a reversed all these changes.Conclusions:Our findings suggest that miR-26a could be a promising therapeutic target for the treatment of cardiorenal injury in CKD.This is attributed to its ability to regulate the LIMS1/ILK signaling pathway,which represents a common molecular mechanism in both heart and kidney tissues.
基金the Ethics Committee of University-Town Hospital of Chongqing Medical University(LL-201682)the Laboratory Animal Ethics Committee of of University-Town Hospital of Chongqing Medical University(2022-053).
文摘Background:Papillary thyroid cancer(PTC)is the most prevalent histological type of differentiated thyroid malignancy.Circular RNAs(circRNAs)have been implicated in the pathogenesis and progression of various cancers.circTIAM1(hsa_circ_0061406)is a novel circRNA with aberrant expression in PTC.However,its functional roles in PTC progression remain to be investigated.Methods:The expression levels of circTIAM1 in the PTC and the matched para-cancerous tissues were detected by quantitative real-time reverse-transcription PCR(qRT-PCR).The subcellular localization of circTIAM1 was examined by fluorescence in-situ hybridization(FISH).Kaplan-Meier plot was used to analyze the association of clinicopathological features with circTIAM1 expression.Bioinformatics databases were utilized to predict the target miRNAs of circTIAM1 and the downstream target mRNAs.RNA pulldown,RIP assay,and dual-luciferase reporter assay were used to confirm the interactions.Functional experiments,such as CCK-8,EDU staining,and apoptosis assays,as well as in vivo xenograft model were employed to explore the impacts of circTIAM1,miR-338-3p,and LIM/SH3 protein 1(LASP1)on the malignant phenotype of the PTC cells.Results:CircTIAM1 was highly expressed in PTC cells.Moreover,circTIAM1 silencing suppressed the proliferation and invasion of PTC cells in vitro and impaired tumorigenesis in vivo.Furthermore,miR-338-3p was verified as a miRNA target of circTIAM1.LASP1 was also identified as a downstream target of miR-338-3p.The anti-tumorigenic effect of miR-338-3p overexpression and the pro-tumorigenic effect of LASP1 was further explored by functional assays,which demonstrated that circTIAM1 modulated the PTC progression through targeting miR-338-3p/LASP1 axis.Conclusion:The overexpression of circTIAM1 is associated with the malignant progression of PTC.A high level of circTIAM1 promotes the malignancy of PTC cells via the miR-338-3p/LASP1 axis.
