目的比较肾移植急性排斥(acute rejection,AR)病人及非AR病人血清miR-663表达水平,在细胞水平上探讨miR-663参与肾移植AR的调控作用,为临床早期诊治AR提供新思路。方法 Real time PCR检测肾移植AR病人及非AR病人血清miR-663表达水平。设...目的比较肾移植急性排斥(acute rejection,AR)病人及非AR病人血清miR-663表达水平,在细胞水平上探讨miR-663参与肾移植AR的调控作用,为临床早期诊治AR提供新思路。方法 Real time PCR检测肾移植AR病人及非AR病人血清miR-663表达水平。设置miR-663 mimic组、miR-663 inhibitor组、阴性对照组及空白对照组;MTT及Annexin V-FITC分别检测过表达miR-663和抑制miR-663表达对人肾小球内皮细胞(HRGEC)生存率和凋亡率的影响;ELISA检测过表达miR-663和抑制miR-663表达对IL-6、IFN-γ、CCL-2及TNF-α表达水平的影响;Transwell实验检测过表达miR-663和抑制miR-663表达对巨噬细胞趋化性的影响。结果 AR组患者血清miR-663表达水平明显较非AR组的肾移植患者升高(4.73±0.28 vs 1.06±0.04;P<0.01)。MTT显示过表达miR-663可以降低HRGEC的生存率并且明显增加其凋亡率,而抑制miR-663则可以降低HRGEC凋亡。过表达miR-663可以明显提高相关炎症因子的表达,同时明显增加巨噬细胞的趋化性。结论 miR-663在肾移植AR过程中发挥着重要作用,可做为早期诊断AR的外周血标志物,并有望成为治疗肾移植AR的一个潜在分子靶点。展开更多
BACKGROUND Recent studies have proved the important role of many oncogenic long noncoding RNAs(lncRNAs) in the progression of pancreatic cancer, but little is known about the mechanisms of tumor suppression in pancrea...BACKGROUND Recent studies have proved the important role of many oncogenic long noncoding RNAs(lncRNAs) in the progression of pancreatic cancer, but little is known about the mechanisms of tumor suppression in pancreatic cancer.AIM To evaluate the function of tumor suppressor lncRNA C9orf139 in pancreatic cancer progression and to study the underlying mechanism.METHODS We assigned 54 patients with pancreatic ductal adenocarcinoma treated at our hospital to the patient group and 30 normal subjects undergoing physical examination to the control group. RT-qPCR was used to measure the relative expression of C9orf139 in the tissue and serum of patients, in an attempt to investigate the prognostic value of C9orf139 in pancreatic cancer patients. The luciferase reporter gene assay was performed to determine the interaction between C9orf139 and miR-663 a. The biological function of C9orf139 was assessed by in vitro assays and in vivo subcutaneous tumor formation tests in animal models. To figure out the molecular mechanism of C9orf139 to act on miR-663 a/Sox12, RNA pull-down, Western blot assay, RNA immunoprecipitation assay, and co-immunoprecipitation assay were performed.RESULTS C9orf139 level significantly increased in the tissue and serum of patients, which had clinical diagnostic value for pancreatic cancer. Patients with high C9orf139 expression had a higher risk of progressing to stage Ⅲ + Ⅳ, lymph node metastasis, and poor differentiation. Cox regression analysis suggested that C9orf139, tumor-node-metastasis stage, and lymph node metastasis were independent prognostic factors in patients. The underlying mechanism of C9orf139 was that it promoted the growth of pancreatic cancer cells by modulating the miR-663 a/Sox12 axis.CONCLUSION C9orf139 is highly expressed in pancreatic cancer, qualified to be used as a potential diagnostic and prognostic marker for pancreatic cancer. Its promotion of pancreatic cancer cell growth is achieved by mediating the miR-663 a/Sox12 axis.展开更多
Objective miR-663 a has been reported to be downregulated by X-ray irradiation and participates in radiation-induced bystander effect via TGF-β1.The goal of this study was to explore the role of mi R-663 a during rad...Objective miR-663 a has been reported to be downregulated by X-ray irradiation and participates in radiation-induced bystander effect via TGF-β1.The goal of this study was to explore the role of mi R-663 a during radiation-induced Epithelium-to-mesenchymal transition(EMT).Methods TGF-β1 or IR was used to induce EMT.After mi R-663 a transfection,cell migration and cell morphological changes were detected and the expression levels of mi R-663 a,TGF-β1,and EMT-related factors were quantified.Results Enhancement of cell migration and promotion of mesenchymal changes induced by either TGF-β1 or radiation were suppressed by mi R-663 a.Furthermore,both X-ray and carbon ion irradiation resulted in the upregulation of TGF-β1 and downregulation of mi R-663 a,while the silencing of TGF-β1 by mi R-663 a reversed the EMT process after radiation.Conclusion Our findings demonstrate an EMT-suppressing effect by mi R-663 a via TGF-β1 in radiationinduced EMT.展开更多
Background:Amphibian-derived peptides exhibit considerable potential in the discovery and development of new therapeutic interventions for clinically challenging chronic skin wounds.MicroRNAs(miRNAs)are also considere...Background:Amphibian-derived peptides exhibit considerable potential in the discovery and development of new therapeutic interventions for clinically challenging chronic skin wounds.MicroRNAs(miRNAs)are also considered promising targets for the development of effective therapies against skin wounds.However,further research in this field is anticipated.This study aims to identify and provide a new peptide drug candidate,as well as to explore the underlying miRNA mechanisms and possible miRNA drug target for skin wound healing.Methods:A combination of Edman degradation,mass spectrometry and cDNA cloning were adopted to determine the amino acid sequence of a peptide thatwas fractionated from the secretion of Odorrana andersonii frog skin using gel-filtration and reversed-phase high-performance liquid chromatography.The toxicity of the peptide was evaluated by Calcein-AM/propidium iodide(PI)double staining against human keratinocytes(HaCaT cells),hemolytic activity against mice blood cells and acute toxicity against mice.The stability of the peptide in plasma was also evaluated.The prohealing potency of the peptide was determined by MTS,scratch healing and a Transwell experiment against HaCaT cells,full-thickness injury wounds and scald wounds in the dorsal skin of mice.miRNA transcriptome sequencing analysis,enzyme-linked immunosorbent assay,real-time polymerase chain reaction and western blotting were performed to explore the molecular mechanisms.Results:A novel peptide homodimer(named OA-GL17d)that contains a disulfide bond between the 16th cysteine residue of the peptide monomer and the sequence‘GLFKWHPRCGEEQSMWT’was identified.Analysis showed that OA-GL17d exhibited no hemolytic activity or acute toxicity,but effectively promoted keratinocyte proliferation and migration and strongly stimulated the repair of full-thickness injury wounds and scald wounds in the dorsal skin of mice.Mechanistically,OA-GL17d decreased the level of miR-663a to increase the level of transforming growth factor-β1(TGF-β1)and activate the subsequent TGF-β1/Smad signaling pathway,thereby resulting in accelerated skin wound re-epithelialization and granular tissue formation.Conclusions:Our results suggest that OA-GL17d is a new peptide drug candidate for skin wound repair.This study emphasizes the importance of exogenous peptides as molecular probes for exploring competing endogenous RNA mechanisms and indicates that miR-663a may be an effective target for promoting skin repair.展开更多
文摘目的比较肾移植急性排斥(acute rejection,AR)病人及非AR病人血清miR-663表达水平,在细胞水平上探讨miR-663参与肾移植AR的调控作用,为临床早期诊治AR提供新思路。方法 Real time PCR检测肾移植AR病人及非AR病人血清miR-663表达水平。设置miR-663 mimic组、miR-663 inhibitor组、阴性对照组及空白对照组;MTT及Annexin V-FITC分别检测过表达miR-663和抑制miR-663表达对人肾小球内皮细胞(HRGEC)生存率和凋亡率的影响;ELISA检测过表达miR-663和抑制miR-663表达对IL-6、IFN-γ、CCL-2及TNF-α表达水平的影响;Transwell实验检测过表达miR-663和抑制miR-663表达对巨噬细胞趋化性的影响。结果 AR组患者血清miR-663表达水平明显较非AR组的肾移植患者升高(4.73±0.28 vs 1.06±0.04;P<0.01)。MTT显示过表达miR-663可以降低HRGEC的生存率并且明显增加其凋亡率,而抑制miR-663则可以降低HRGEC凋亡。过表达miR-663可以明显提高相关炎症因子的表达,同时明显增加巨噬细胞的趋化性。结论 miR-663在肾移植AR过程中发挥着重要作用,可做为早期诊断AR的外周血标志物,并有望成为治疗肾移植AR的一个潜在分子靶点。
文摘BACKGROUND Recent studies have proved the important role of many oncogenic long noncoding RNAs(lncRNAs) in the progression of pancreatic cancer, but little is known about the mechanisms of tumor suppression in pancreatic cancer.AIM To evaluate the function of tumor suppressor lncRNA C9orf139 in pancreatic cancer progression and to study the underlying mechanism.METHODS We assigned 54 patients with pancreatic ductal adenocarcinoma treated at our hospital to the patient group and 30 normal subjects undergoing physical examination to the control group. RT-qPCR was used to measure the relative expression of C9orf139 in the tissue and serum of patients, in an attempt to investigate the prognostic value of C9orf139 in pancreatic cancer patients. The luciferase reporter gene assay was performed to determine the interaction between C9orf139 and miR-663 a. The biological function of C9orf139 was assessed by in vitro assays and in vivo subcutaneous tumor formation tests in animal models. To figure out the molecular mechanism of C9orf139 to act on miR-663 a/Sox12, RNA pull-down, Western blot assay, RNA immunoprecipitation assay, and co-immunoprecipitation assay were performed.RESULTS C9orf139 level significantly increased in the tissue and serum of patients, which had clinical diagnostic value for pancreatic cancer. Patients with high C9orf139 expression had a higher risk of progressing to stage Ⅲ + Ⅳ, lymph node metastasis, and poor differentiation. Cox regression analysis suggested that C9orf139, tumor-node-metastasis stage, and lymph node metastasis were independent prognostic factors in patients. The underlying mechanism of C9orf139 was that it promoted the growth of pancreatic cancer cells by modulating the miR-663 a/Sox12 axis.CONCLUSION C9orf139 is highly expressed in pancreatic cancer, qualified to be used as a potential diagnostic and prognostic marker for pancreatic cancer. Its promotion of pancreatic cancer cell growth is achieved by mediating the miR-663 a/Sox12 axis.
基金funded by the National Natural Science Foundation of China[no.U1932208,YANG Kai]the Science and Technology Research Project of Gansu Province[no.17JR5RA307 and 145RTSA012,WANG Ju Fang]the Science and Technology Research Project of Gansu Province[no.21JR7RA108,DING Nan]
文摘Objective miR-663 a has been reported to be downregulated by X-ray irradiation and participates in radiation-induced bystander effect via TGF-β1.The goal of this study was to explore the role of mi R-663 a during radiation-induced Epithelium-to-mesenchymal transition(EMT).Methods TGF-β1 or IR was used to induce EMT.After mi R-663 a transfection,cell migration and cell morphological changes were detected and the expression levels of mi R-663 a,TGF-β1,and EMT-related factors were quantified.Results Enhancement of cell migration and promotion of mesenchymal changes induced by either TGF-β1 or radiation were suppressed by mi R-663 a.Furthermore,both X-ray and carbon ion irradiation resulted in the upregulation of TGF-β1 and downregulation of mi R-663 a,while the silencing of TGF-β1 by mi R-663 a reversed the EMT process after radiation.Conclusion Our findings demonstrate an EMT-suppressing effect by mi R-663 a via TGF-β1 in radiationinduced EMT.
基金supported by grants from the National Natural Science Foundation of China(81760648,32060212 and 82160159)Yunnan Applied Basic Research Project Foundation(2019FB128)+4 种基金Project of Yunnan Applied Basic Research Project-Kunming Medical University Union Foundation(202101AY070001-006 and 2019FE001(-183))Program for Innovative Research Team in Ministry of Education of China(IRT17-R49)Science and Technology Leadership Talent Project in Yunnan China(2017HA010)Endocrine Clinical Medical Center of Yunnan Province(ZX2019-02-02)the Innovative Team of Precise Prevention and Treatment against Metabolic Diseases of Yunnan University,Scientific Research Fund Projects from the Department of Education of Yunnan Province(2021 J0205).
文摘Background:Amphibian-derived peptides exhibit considerable potential in the discovery and development of new therapeutic interventions for clinically challenging chronic skin wounds.MicroRNAs(miRNAs)are also considered promising targets for the development of effective therapies against skin wounds.However,further research in this field is anticipated.This study aims to identify and provide a new peptide drug candidate,as well as to explore the underlying miRNA mechanisms and possible miRNA drug target for skin wound healing.Methods:A combination of Edman degradation,mass spectrometry and cDNA cloning were adopted to determine the amino acid sequence of a peptide thatwas fractionated from the secretion of Odorrana andersonii frog skin using gel-filtration and reversed-phase high-performance liquid chromatography.The toxicity of the peptide was evaluated by Calcein-AM/propidium iodide(PI)double staining against human keratinocytes(HaCaT cells),hemolytic activity against mice blood cells and acute toxicity against mice.The stability of the peptide in plasma was also evaluated.The prohealing potency of the peptide was determined by MTS,scratch healing and a Transwell experiment against HaCaT cells,full-thickness injury wounds and scald wounds in the dorsal skin of mice.miRNA transcriptome sequencing analysis,enzyme-linked immunosorbent assay,real-time polymerase chain reaction and western blotting were performed to explore the molecular mechanisms.Results:A novel peptide homodimer(named OA-GL17d)that contains a disulfide bond between the 16th cysteine residue of the peptide monomer and the sequence‘GLFKWHPRCGEEQSMWT’was identified.Analysis showed that OA-GL17d exhibited no hemolytic activity or acute toxicity,but effectively promoted keratinocyte proliferation and migration and strongly stimulated the repair of full-thickness injury wounds and scald wounds in the dorsal skin of mice.Mechanistically,OA-GL17d decreased the level of miR-663a to increase the level of transforming growth factor-β1(TGF-β1)and activate the subsequent TGF-β1/Smad signaling pathway,thereby resulting in accelerated skin wound re-epithelialization and granular tissue formation.Conclusions:Our results suggest that OA-GL17d is a new peptide drug candidate for skin wound repair.This study emphasizes the importance of exogenous peptides as molecular probes for exploring competing endogenous RNA mechanisms and indicates that miR-663a may be an effective target for promoting skin repair.
