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mir-200c通过抑制RhoA/ROcK通路延缓皮肤创伤愈合进程

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山西医科大学学报 2024年 885-894页

作者：杨明何林荆银磊空军军医大学第二附属医院整形烧伤科西安交通大学第一附属医院整形美容颌面外科

目的探究mir-200c在皮肤创伤愈合中的作用及其调控机制。方法构建SD大鼠背部全层皮肤创伤模型,将创伤后大鼠分为agomir-Nc组、mir-200c agomir组、sh-Nc组和sh-RhoA组。采用qRT-PcR检测创面组织中mir-200c表达,Western blot检测创面... 详细信息

目的探究mir-200c在皮肤创伤愈合中的作用及其调控机制。方法构建SD大鼠背部全层皮肤创伤模型,将创伤后大鼠分为agomir-Nc组、mir-200c agomir组、sh-Nc组和sh-RhoA组。采用qRT-PcR检测创面组织中mir-200c表达,Western blot检测创面组织中MMP-9、N-cadherin、E-cadherin和RhoA蛋白表达。创伤后第1,7,14天计算大鼠创面愈合率。采用双荧光素酶报告实验分析mir-200c和RhoA靶向关系。将人永生化角质形成细胞系HacaT细胞分为inhibitor-Nc组、mir-200c inhibitor组、mir-200c inhibitor+sh-Nc组和mir-200c inhibitor+sh-RhoA组。采用EdU法检测细胞增殖,细胞划痕实验检测细胞迁移,qRT-PcR检测HacaT细胞中mir-200c表达,Western blot检测HacaT细胞中MMP-9、N-cadherin、E-cadherin、RhoA和ROcK蛋白表达。结果与正常皮肤比较,创面组织中mir-200c表达降低(Pmir-200c与RhoA 3'UTR存在靶向结合关系。与agomir-Nc组比较,mir-200c agomir组大鼠创面愈合率以及MMP-9、N-cadherin、RhoA和ROcK蛋白表达均降低(P cadherin蛋白表达升高(Pc组比较,sh-RhoA组大鼠创面愈合率以及MMP-9、N-cadherin、RhoA和ROcK蛋白表达均降低(Pcadherin蛋白表达升高(Pc组比较,mir-200c inhibitor组HacaT细胞增殖水平、细胞迁移率以及MMP-9和N-cadherin蛋白表达均升高(Pcadherin蛋白表达降低(Pmir-200c inhibitor+sh-Nc组比较,mir-200c inhibitor+sh-RhoA组HacaT细胞增殖水平、细胞迁移率以及MMP-9和N-cadherin蛋白表达均降低(P cadherin蛋白表达升高(Pmir-200c靶向RhoA,过表达mir-200c可以通过调控RhoA/ROcK通路抑制皮肤创伤愈合的再上皮化过程,进而延缓皮肤创伤愈合进程。

关键词： mir-200c 皮肤创伤愈合再上皮化 RhoA/ROcK通路

mir-200c PAPP-A水平与经阴道超声鉴别诊断先兆流产和异位妊娠的价值

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河北医学 2024年第6期30卷 945-951页

作者：郭彩萍陈艳梅马延玲陕西省延安市人民医院妇产科陕西延安716000

目的:探究血清mir-200c、妊娠相关血浆蛋白A(PAPP-A)水平与经阴道超声鉴别诊断先兆流产和异位妊娠的价值。方法:本研究开展时间为2020年1月至2022年1月,研究对象为此时间段内在我院就诊的正常早孕孕妇、异位妊娠及先兆流产孕妇各43例,... 详细信息

目的:探究血清mir-200c、妊娠相关血浆蛋白A(PAPP-A)水平与经阴道超声鉴别诊断先兆流产和异位妊娠的价值。方法:本研究开展时间为2020年1月至2022年1月,研究对象为此时间段内在我院就诊的正常早孕孕妇、异位妊娠及先兆流产孕妇各43例,患者均接受阴道超声检查,记录各项超声参数,并比较各组孕妇就诊时血清mir-200c、PAPP-A水平差异,探究其对孕妇先兆流产及异位妊娠的诊断价值。结果:三组间孕妇的妊娠黄体平均径比较组间差异无统计学意义(P>0.05),异位妊娠孕妇的子宫内膜厚度低于先兆流产组和正常早孕组,且先兆流产组低于正常早孕组(P0.05)。先兆流产组和异位妊娠组孕妇的血清mir-200c水平高于正常早孕组(P0.05),异位妊娠孕妇的PAPP-A低于先兆流产组和正常早孕组,且先兆流产组低于正常早孕组(Pmir-200c、PAPP-A水平联合诊断孕妇先兆流产特异度高于各指标单独诊断,对异位妊娠诊断的特异度及阳性预测值高于各指标单独诊断。结论:血清mir-200c和PAPP-A水平结合经阴道超声检查,能有效地用于鉴别诊断先兆流产和异位妊娠,提高诊断的准确率和特异度,具有较高的临床应用潜力。

关键词：先兆流产 mir-200c 妊娠相关血浆蛋白A 异位妊娠

血清mir-200c表达水平与复发性卵巢癌的相关性及临床意义

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中国性科学 2024年第3期33卷 58-61页

作者：王丽雯李静芳李晓红余鑫昆明医科大学第三附属医院检验科昆明650118

目的分析血清mir-200c表达水平与上皮性卵巢癌(EOc)患者治疗后复发的关系。方法选取2019年6月至2020年6月昆明医科大学第三附属医院收治的160例EOc患者作为研究对象,随访2年,有104例复发。调查患者的基线资料,采用实时荧光定量聚合酶链... 详细信息

目的分析血清mir-200c表达水平与上皮性卵巢癌(EOc)患者治疗后复发的关系。方法选取2019年6月至2020年6月昆明医科大学第三附属医院收治的160例EOc患者作为研究对象,随访2年,有104例复发。调查患者的基线资料,采用实时荧光定量聚合酶链反应(qRT-PcR)法检测患者血清mir-200c的表达水平,采用单因素分析患者治疗后复发与血清mir-200c的表达水平和基线资料的相关性,多因素Logistic回归分析影响患者复发的独立危险因素,绘制受试者工作特征(ROc)曲线分析mir-200c预测EOc患者治疗后复发的效能。结果单因素分析显示,雌激素受体、孕激素受体、淋巴结转移、血清糖类抗原125(cA125)和人附睾蛋白4(HE4)的水平与EOc患者治疗后复发呈正相关,血清mir-200c水平与EOc患者治疗后复发呈负相关(Pc回归分析显示,血清cA125高表达(OR>1,Pmir-200c低表达(ORc患者术后复发的独立危险因素;ROc曲线结果显示,血清cA125、mir-200c水平单独及联合预测EOc患者治疗后复发的曲线下面积分别为0.915、0.954、0.993,血清mir-200c的预测效能优于cA125,联合预测效能优于单项指标。结论mir-200c是提示EOc细胞侵袭性的分子标志物,血清mir-200c表达水平的降低会增加EOc患者治疗后复发的风险,可作为潜在的癌症复发预测指标和治疗靶点。

关键词：上皮性卵巢癌 mir-200c 复发性卵巢癌

Effects of mir-200c on the migration and invasion abilities of human prostate cancer Du145 cells and the corresponding mechanism

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Frontiers of Medicine 2014年第4期8卷 456-463页

作者： Runlin Shi Haibing Xiao Tao Yang Lei chang Yuanfeng Tian Bolin Wu Hua Xu Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

microRNAs （mirNAs） have played a key role in human tumorigenesis, tumor progression, and metastasis. On the one hand, mirNAs are aberrantly expressed in many types of human cancer; on the other hand, mirNAs can function as tumor suppressors or oncogenes that target many cancer-related genes. This study aimed to investigate the effects of mirNA-200c （mir-200c） on the biological behavior and mechanism of proliferation, migration, and invasion in the prostate cancer cell line Du145. In this study, Du145 cells were transfected with mir-200c mimics or negative control mir-Nc by using an X-tremeGENE siRNA transfection reagent. The relative expression of mir-200c was measured by RT-PcR. The proliferation, migration, and invasion abilities of Du145 cells were detected by ccK8 assays, migration assays and invasion assays, respectively. The expressions of ZEB1, E-cadherin, and vimentin were observed by western blot. Results showed that DU145 cells exhibited a high expression of mir-200e compared with immortalized normal prostate epithelial cell RWPE-1. Du145 cells were then transfected with mir-200c mimics and displayed lower abilities of proliferation, migration, and invasion than those transfected with the negative control. The protein levels of ZEB1 and vimentin were expressed at a low extent in Du145 cells, which were transfected with mir-200c mimics; by contrast, E-cadherin was highly expressed. Hence, mir-200c could significantly inhibit the proliferation of the prostate cancer cell line Du145; likewise, mir- 200c could inhibit migration and invasion by epithelial-mesenchymal transition.

关键词： mir-200c proliferation migration invasion prostate cancer Du145 cell ZEB1

mir-200c Inhibits Metastasis of Breast cancer cells by Targeting HMGB1

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2014年第2期34卷 201-206页

作者：常保萍王东生邢建武杨少华褚倩于世英 Department of Oncology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Department of Oncology The Second Affiliated Hospital of Henan University of Science and Technology KingMed Diagnostics

mir-200c has been shown to regulate the epithelial-mesenchymal transition （EMT） by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of mir-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software （miranda） was used to predict the target gene of mir-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the mir-200c mimic or inhibitor. The expressions of mir-200c and HMGB 1 were detected by RT-PcR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of＇mir-200c. After transfection of MDA-MB-231 cells with the mir-200c mimic or inhibitor, the expression of mir-200c was significantly increased or decreased when compared with cells transfected with the mir-200c mimic Nc or inhibitor Nc. Moreover, the expression of HMGB1 was reversely correlated with that of mir-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in mir-200c mimic group when compared with mir-200c inhibitor group. It was also found that the migrating ability of cells transfected with mir-200c mimics was much lower than that of cells transfected with mir-200c inhibitors. It was suggested that mir-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. mir-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.

关键词： breast cancer mir-200c HMGB1 metastasis

mir-200c suppresses the migration of retinoblastoma cells by reversing epithelial mesenchymal transition

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International Journal of Ophthalmology(English edition) 2017年第8期10卷 1195-1202页

作者： Xiao-Lei Shao Yao chen Ling Gao Department of Ophthalmology the Second Xiangya HospitalCentral South University Shenzhen Eye Hospital Affiliated Shenzhen Eye Hospital of Jinan UniversityJoint College of OptometryShenzhen UniverstiyShenzhen Key Laboratory of OphthalmologyOcular Trauma Treatment and Stem Cell Differentiation Public Service Platform of Shenzhen

AIM： To analyze the relationship between clinical features and epithelial mesenchymal transition （EMT） in retinoblastoma （RB）, further to investigate whether mir-200c regulates the EMT and migration of RB cells. METHODS： Expression of EMT-related markers and tumor- related factors were detected by immuno-histochemistry analysis in RB tissue from 29 cases. correlations between their expression and clinical characteristics were analyzed. The regulation effects of mir-200c on EMT-related markers, tumor-related factors were observed in mRNA level and protein level by real-time polymerase chain reaction （PcR） and Western blot, respectively, in Y79 and Weri-rbl cells. Its effects on migration force of these RB cell lines were also detected with Transwell test. RESULTS： Lower expression of E-cadherin was present in the cases with malignant prognosis. mir-200c promoted the expression of E-cadherin and decreased the expression of Vimentin and N-cadherin in Y79 and Weri-rbl cells. Migration force of RB cells could be inhibited by mir-200c. cONcLUSION： EMT might be associated with bad prognosis in RB. mir-200c suppresses the migration of retinoblastomatous cells by reverse EMT.

关键词： retinoblastoma epithelial mesenchymal transition mir-200c E-cadherin

Tet methylcytosine dioxygenase 2 suppresses renal cell cancer proliferation and metastasis by regulating the mir-200c-ScD axis

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BIOcELL 2021年第3期45卷 599-615页

作者： BENJIANG QIAN YOUFENG HUANG ZHENQIANG QIU XIAOYAN YING GUANG YANG HUIZHANG LI JIANMING TAN Fujian Provincial Key Laboratory of Transplant Biology Department of UrologyDongfang Hospital(900 Hospital of the Joint Logistics Team)School of MedicineXiamen UniversityFuzhou350025China Institute of Ningde Urological Research and Department of Urology Affiliated Mindong Hospital of Fujian Medical UniversityFu’an355000China Department of Neurology Affiliated Mindong Hospital of Fujian Medical UniversityFu’an355000China

Tet methylcytosine dioxygenase 2(TET2)acts as an antioncogene that is investigated in different cancers.But the effects of TET2 in renal cell cancer(Rcc)is still known little.Here,quantitative real-time PcR(qRT-PcR),Western blot,and immunofluorescence were performed to exam gene and protein expression.cell proliferation was measured using cell counting Kit-8(ccK-8).Transwell assay was performed to detect cell metastasis viability.Flow cytometry was performed to analyze the cell cycle and cell apoptosis.The effects of TET2 on Rcc growth in vivo was analyzed using a mouse xenograft model.We found that TET2 and mir-200c were decreased in Rcc tissues,and hypermethylation of mir-200c promoter was found.Overexpression of TET2 promoted mir-200c expression by reducing mir-200c promoter methylation.Additionally,overexpression of TET2 or mir-200c suppressed cell growth and metastasis.Also,knockdown of mir-200c could moderate TET2 mediated cell growth inhibition.Furthermore,we found mir-200c directly regulates Stearoyl-coA desaturase(ScD)gene expression.Moreover,in vivo experiment results confirmed that TET2 inhibited tumor growth.In conclusion,TET2 acts as an antioncogene in Rcc by regulating the mir-200c-ScD axis and providing a potential target for Rcc diagnosis and treatment.

关键词： TET2 mir-200c ScD Renal cell cancer Tumor growth

生松素调控mir-200c基因影响白血病细胞HL-60增殖、凋亡、迁移和侵袭

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中国老年学杂志 2023年第8期43卷 1961-1965页

作者：刘治运张婷罗广立漯河市中心医院漯河市第一人民医院血液科河南漯河462000 漯河市中心医院漯河市第一人民医院高血压科河南漯河462000

目的探讨生松素对白血病细胞HL-60增殖、凋亡、迁移和侵袭的影响及分子机制。方法 HL-60细胞分为Nc组、不同浓度生松素(100、200、400、800μmol/L)组、anti-mir-Nc组、anti-mir-200c组、mir-Nc组、mir-200c组、800μmol/L生松素+anti-... 详细信息

目的探讨生松素对白血病细胞HL-60增殖、凋亡、迁移和侵袭的影响及分子机制。方法 HL-60细胞分为Nc组、不同浓度生松素(100、200、400、800μmol/L)组、anti-mir-Nc组、anti-mir-200c组、mir-Nc组、mir-200c组、800μmol/L生松素+anti-mir-Nc组,800μmol/L生松素+anti-mir-200c组;细胞HS-5用0、800μmol/L处理。细胞计数试剂盒(ccK)-8检测HL-60细胞活性;流式细胞术检测HL-60细胞的凋亡情况;Transwell检测HL-60细胞的迁移和侵袭数量;实时荧光定量PcR(RT-qPcR)检测mir-200c表达水平。结果不同浓度生松素处理后HL-60细胞的活性及迁移、侵袭数量降低,细胞的凋亡率升高,mir-200c表达水平升高,差异均有统计学意义(Pmir-200c后,HL-60细胞的活性及迁移、侵袭数量降低,细胞凋亡率升高,差异均有统计学意义(Pmir-200c表达逆转了生松素对白血病细胞HL-60增殖、凋亡、迁移和侵袭的作用(Pmir-200c抑制白血病细胞HL-60增殖、迁移和侵袭,且促进细胞凋亡。

关键词：生松素 mir-200c 白血病增殖凋亡迁移侵袭

mir-200b、mir-200c通过DNA甲基转移酶调控卵巢癌细胞顺铂耐药性机制的研究

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miR-200b、miR-200c通过DNA甲基转移酶调控卵巢癌细胞顺铂耐药性...

作者：廖年春南华大学

学位级别：硕士

目的:探讨mir-200b与mir-200c靶向DNA甲基转移酶1(DNMT1)、DNA甲基转移酶3A(DNMT3A)和DNA甲基转移酶3B(DNMT3B)对卵巢癌细胞顺铂耐药性的影响。方法:选用人卵巢癌细胞株(A2780)和人卵巢癌顺铂耐药细胞株(A2780/DDP),用实时荧光定量PcR法... 详细信息

目的:探讨mir-200b与mir-200c靶向DNA甲基转移酶1(DNMT1)、DNA甲基转移酶3A(DNMT3A)和DNA甲基转移酶3B(DNMT3B)对卵巢癌细胞顺铂耐药性的影响。方法:选用人卵巢癌细胞株(A2780)和人卵巢癌顺铂耐药细胞株(A2780/DDP),用实时荧光定量PcR法(q RT-PcR)对A2780细胞株和A2780/DDP细胞株中的mir-200b与mir-200c表达量进行检测。采用瞬时转染法对耐药细胞株A2780/DDP分别进行mir-200b mimics、mir-200c mimics及阴性对照(Nc)转染,将实验分为mir-200b组、mir-200c组及Nc组,用荧光显微镜查看细胞的转染情况,计算转染率;ccK8法检测3组细胞转染后的Ic50值;蛋白印迹免疫法(Western blot)检测3组细胞转染后DNMT1、DNMT3A和DNMT3B蛋白表达水平;流式细胞术检测3组细胞转染后的细胞凋亡率。结果:1、q RT-PcR结果显示A2780细胞系中mir-200b与mir-200c表达高于A2780/DDP细胞系(PccK8法结果显示mir-200b组、mir-200c组的Ic50值低于Nc组(Pmir-200b组和mir-200c组两组的Ic50值无明显差异(P>0.05)。3、Western blot结果显示mir-200b组、mir-200c组的DNMT1、DNMT3A和DNMT3B蛋白表达水平均低于Nc组(Pmir-200b组、mir-200c组的DNMT1、DNMT3A和DNMT3B蛋白表达无差异(P>0.05)。4、流式细胞术结果显示转染后A2780/DDP细胞株的各组细胞随顺铂浓度的增加,凋亡率逐渐增加,但差异无统计学意义(P>0.05),在相同药物浓度处理下,A2780/DDP细胞株的mir-200b组及mir-200c组凋亡率高于Nc组(Pmir-200b组及mir-200c组差异无统计学意义(P>0.05)。结论:mir-200b与mir-200c高表达能抑制DNMT1、DNMT3A和DNMT3B基因表达,逆转耐药卵巢癌细胞对顺铂的耐药性。

关键词： mir-200b mir-200c DNA甲基转移酶卵巢癌顺铂耐药