目的检测微小核糖核酸(mircro RNAs,miR)-450a-5p和环磷酸腺苷反应元件结合蛋白(c-AMP response element binding protein,CREB)在妊娠期糖尿病(gestational diabetes mellitus,GDM)患者血清中的表达情况,探讨其对GDM诊断的临床意义。...目的检测微小核糖核酸(mircro RNAs,miR)-450a-5p和环磷酸腺苷反应元件结合蛋白(c-AMP response element binding protein,CREB)在妊娠期糖尿病(gestational diabetes mellitus,GDM)患者血清中的表达情况,探讨其对GDM诊断的临床意义。方法选取2018年4月~2020年4月在如皋市人民医院妇产科建档产检的120例GDM孕妇为GDM组,选择同期120例健康孕妇为对照组。采用实时荧光定量PCR(qRT-PCR)法检测受试者血清中miR-450a-5p水平;采用酶联免疫吸附实验(ELISA)检测受试者血清中CREB表达水平;Pearson法分析血清miR-450a-5p和CREB水平与GDM孕妇临床病理特征相关性;受试者工作特征(ROC)曲线分析血清中miR-450a-5p和CREB表达水平对GDM的诊断价值以及采用Logistic回归分析GDM发生的影响因素。结果与对照组相比,GDM组患者血清中miR-450a-5p表达水平(0.69±0.15 vs 1.58±0.34)显著下调(t=26.235,P<0.05),血清CREB表达水平(8.75±2.42μg/ml vs 2.64±0.66μg/ml)显著上调(t=26.683,P<0.05),差异均有统计学意义。血清miR-450a-5p水平与空腹血糖(fasting plasma glucose,FPG)和稳态模型胰岛素抵抗指数(homeostasis model assessment-insulin resistance,HOMA-IR)呈显著负相关(r=-0.427,-0.251;P=0.027,0.015),与空腹胰岛素(fasting insulin,FINS)呈显著正相关(r=0.636,P=0.005);CREB水平与FBG,HOMA-IR呈显著正相关(r=0.432,0.574;P=0.041,0.008),与FINS呈显著负相关(r=-0.573,P=0.028),其差异均具有统计学意义。ROC曲线显示,血清miR-450a-5p和CREB水平对GDM诊断的曲线下面积分别为0.926(敏感度和特异度分别为91.7%和90.8%),0.881(敏感度和特异度分别为86.7%和89.2%)。Logistic回归分析显示miR-450a-5p是GDM的保护因素,血清CREB是GDM的独立危险因素。结论miR-450a-5p和CREB在GDM患者血清中异常表达,且与胰岛素抵抗相关指标有一定的相关性。miR-450a-5p和CREB检测对于GDM临床诊断具有重要参考价值。展开更多
Background:The sensitivity of breast cancer cells to radiation is a key cause of locoregional recurrence after postoperative radiotherapy.Several studies have reported that microRNAs(miRNAs)are involved in the radiose...Background:The sensitivity of breast cancer cells to radiation is a key cause of locoregional recurrence after postoperative radiotherapy.Several studies have reported that microRNAs(miRNAs)are involved in the radiosensitivity of human breast cancer cells.One miRNA microarray study showed that miR-450b-5p was overexpressed 13.3-fold in patients with estrogen receptor–positive(ER^(+))and human epidermal growth factor receptor 2–negative(HER2−)breast cancer and no local relapse compared with local relapse patients.However,its underlying mechanism of action remains unknown.Methods:The predicted target mRNAs of miR-450b-5p were screened using the TargetScan,miRDB,and miRWalk databases.Western blotting,quantitative polymerase chain reaction,and dual-luciferase reporter assays explored the association between cyclindependent kinase 6(CDK6)and miR-450b-5p.The cell counting kit-8 assay and flow cytometry detected the proliferation of transfected MCF7 cells.Colony formation and xenograft tumors detected the radiosensitivity of the transfected MCF7 cells.Results:Bioinformatics analysis,Western blotting,quantitative polymerase chain reaction,and dual-luciferase reporter assays demonstrated that CDK6 was the target gene of miR-450b-5p.Furthermore,in vitro and in vivo experiments showed that miR-450b-5p inhibited MCF7 cell proliferation and cell cycle progression,increased the sensitizer enhancement ratio,and decreased the volume of xenograft tumors after irradiation by regulating CDK6.Conclusions:This study demonstrates that miR-450b-5p enhances the radiosensitivity of hormone receptor–positive(HR^(+))and HER2−breast cancer cells and elucidates its mechanism.miR-450b-5p may be considered a therapeutic target in HR^(+)and HER2−breast cancer treated with radiotherapy.展开更多
目的探讨Gm31083/miR-450b-5p/EphB1通路介导肺癌骨转移癌痛小鼠的机制。方法2020年1月至2021年12月,选取健康SPF级SD雄性小鼠20只,采用随机数字表法分为假手术组、模型组各10只。肺癌骨转移癌痛模型通过股骨内注射人肺腺癌A549细胞建...目的探讨Gm31083/miR-450b-5p/EphB1通路介导肺癌骨转移癌痛小鼠的机制。方法2020年1月至2021年12月,选取健康SPF级SD雄性小鼠20只,采用随机数字表法分为假手术组、模型组各10只。肺癌骨转移癌痛模型通过股骨内注射人肺腺癌A549细胞建立。于小鼠左侧足底中部采用2 g Von Frey细纤维刺激,5~6 s观察小鼠缩足反应;于热板仪上测定小鼠的痛阈值;采用实时荧光定量聚合酶链式反应法测定miR-450b-5p表达;采用Western Blot测定Gm31083和EphB1蛋白表达。结果模型组小鼠缩足反应率[(31.98±6.36)%]高于假手术组[(22.78±4.54)%](t=-3.72,P<0.05)。模型组小鼠热刺激痛阈值[(8.18±2.53)s]低于假手术组[(15.42±3.97)s](t=4.86,P<0.001)。模型组小鼠miR-450b-5p相对表达量(1.62±0.29)高于假手术组(1.00±0.04)(t=-6.70,P<0.001)。模型组小鼠Gm31083、EphB1蛋白表达灰度值分别为(1.23±0.21)、(2.73±0.28),均高于假手术组[(0.67±0.18)、(1.25±0.24)](t=-6.40、-12.69,均P<0.001)。结论Gm31083/miR-450b-5p/EphB1通路在肺癌骨转移癌痛小鼠中高表达,有可能成为评估肺癌骨转移癌痛新的标志物。展开更多
文摘目的检测微小核糖核酸(mircro RNAs,miR)-450a-5p和环磷酸腺苷反应元件结合蛋白(c-AMP response element binding protein,CREB)在妊娠期糖尿病(gestational diabetes mellitus,GDM)患者血清中的表达情况,探讨其对GDM诊断的临床意义。方法选取2018年4月~2020年4月在如皋市人民医院妇产科建档产检的120例GDM孕妇为GDM组,选择同期120例健康孕妇为对照组。采用实时荧光定量PCR(qRT-PCR)法检测受试者血清中miR-450a-5p水平;采用酶联免疫吸附实验(ELISA)检测受试者血清中CREB表达水平;Pearson法分析血清miR-450a-5p和CREB水平与GDM孕妇临床病理特征相关性;受试者工作特征(ROC)曲线分析血清中miR-450a-5p和CREB表达水平对GDM的诊断价值以及采用Logistic回归分析GDM发生的影响因素。结果与对照组相比,GDM组患者血清中miR-450a-5p表达水平(0.69±0.15 vs 1.58±0.34)显著下调(t=26.235,P<0.05),血清CREB表达水平(8.75±2.42μg/ml vs 2.64±0.66μg/ml)显著上调(t=26.683,P<0.05),差异均有统计学意义。血清miR-450a-5p水平与空腹血糖(fasting plasma glucose,FPG)和稳态模型胰岛素抵抗指数(homeostasis model assessment-insulin resistance,HOMA-IR)呈显著负相关(r=-0.427,-0.251;P=0.027,0.015),与空腹胰岛素(fasting insulin,FINS)呈显著正相关(r=0.636,P=0.005);CREB水平与FBG,HOMA-IR呈显著正相关(r=0.432,0.574;P=0.041,0.008),与FINS呈显著负相关(r=-0.573,P=0.028),其差异均具有统计学意义。ROC曲线显示,血清miR-450a-5p和CREB水平对GDM诊断的曲线下面积分别为0.926(敏感度和特异度分别为91.7%和90.8%),0.881(敏感度和特异度分别为86.7%和89.2%)。Logistic回归分析显示miR-450a-5p是GDM的保护因素,血清CREB是GDM的独立危险因素。结论miR-450a-5p和CREB在GDM患者血清中异常表达,且与胰岛素抵抗相关指标有一定的相关性。miR-450a-5p和CREB检测对于GDM临床诊断具有重要参考价值。
文摘Background:The sensitivity of breast cancer cells to radiation is a key cause of locoregional recurrence after postoperative radiotherapy.Several studies have reported that microRNAs(miRNAs)are involved in the radiosensitivity of human breast cancer cells.One miRNA microarray study showed that miR-450b-5p was overexpressed 13.3-fold in patients with estrogen receptor–positive(ER^(+))and human epidermal growth factor receptor 2–negative(HER2−)breast cancer and no local relapse compared with local relapse patients.However,its underlying mechanism of action remains unknown.Methods:The predicted target mRNAs of miR-450b-5p were screened using the TargetScan,miRDB,and miRWalk databases.Western blotting,quantitative polymerase chain reaction,and dual-luciferase reporter assays explored the association between cyclindependent kinase 6(CDK6)and miR-450b-5p.The cell counting kit-8 assay and flow cytometry detected the proliferation of transfected MCF7 cells.Colony formation and xenograft tumors detected the radiosensitivity of the transfected MCF7 cells.Results:Bioinformatics analysis,Western blotting,quantitative polymerase chain reaction,and dual-luciferase reporter assays demonstrated that CDK6 was the target gene of miR-450b-5p.Furthermore,in vitro and in vivo experiments showed that miR-450b-5p inhibited MCF7 cell proliferation and cell cycle progression,increased the sensitizer enhancement ratio,and decreased the volume of xenograft tumors after irradiation by regulating CDK6.Conclusions:This study demonstrates that miR-450b-5p enhances the radiosensitivity of hormone receptor–positive(HR^(+))and HER2−breast cancer cells and elucidates its mechanism.miR-450b-5p may be considered a therapeutic target in HR^(+)and HER2−breast cancer treated with radiotherapy.
文摘目的探讨Gm31083/miR-450b-5p/EphB1通路介导肺癌骨转移癌痛小鼠的机制。方法2020年1月至2021年12月,选取健康SPF级SD雄性小鼠20只,采用随机数字表法分为假手术组、模型组各10只。肺癌骨转移癌痛模型通过股骨内注射人肺腺癌A549细胞建立。于小鼠左侧足底中部采用2 g Von Frey细纤维刺激,5~6 s观察小鼠缩足反应;于热板仪上测定小鼠的痛阈值;采用实时荧光定量聚合酶链式反应法测定miR-450b-5p表达;采用Western Blot测定Gm31083和EphB1蛋白表达。结果模型组小鼠缩足反应率[(31.98±6.36)%]高于假手术组[(22.78±4.54)%](t=-3.72,P<0.05)。模型组小鼠热刺激痛阈值[(8.18±2.53)s]低于假手术组[(15.42±3.97)s](t=4.86,P<0.001)。模型组小鼠miR-450b-5p相对表达量(1.62±0.29)高于假手术组(1.00±0.04)(t=-6.70,P<0.001)。模型组小鼠Gm31083、EphB1蛋白表达灰度值分别为(1.23±0.21)、(2.73±0.28),均高于假手术组[(0.67±0.18)、(1.25±0.24)](t=-6.40、-12.69,均P<0.001)。结论Gm31083/miR-450b-5p/EphB1通路在肺癌骨转移癌痛小鼠中高表达,有可能成为评估肺癌骨转移癌痛新的标志物。
