目的探讨微小RNA-6321(microRNA-6321,miR-6321)对大鼠心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)的作用,并进一步分析其作用机制与N6-甲基腺苷甲基化之间的关系。方法随机从36只雄性SD大鼠中取12只,分为预实验...目的探讨微小RNA-6321(microRNA-6321,miR-6321)对大鼠心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)的作用,并进一步分析其作用机制与N6-甲基腺苷甲基化之间的关系。方法随机从36只雄性SD大鼠中取12只,分为预实验假手术组和预实验模型组,每组6只,用于验证MIRI模型。预实验假手术组在左前降支下方穿线而不结扎,预实验模型组结扎左前降支30 min后解开结扎线再灌注2 h。随后,本研究将剩余24只大鼠随机分为4组:假手术组、模型组(MIRI组)、腺相关病毒阴性对照组(adeno-associated virus negative control group,AAV-NC组)以及miR-6321过表达干预组(AAV-miR-6321组),每组6只。AAV-NC组和AAV-miR-6321组在模型建立前两周经尾静脉单次注射AAV-NC和AAV-miR-6321。采用冠状动脉左前降支结扎法建立大鼠MIRI模型。假手术组在左前降支下方穿线而不结扎,MIRI组、AAV-NC组和AAV-miR-6321组均进行缺血30 min、再灌注120 min处理。采用苏木精-伊红染色检测分析心肌组织损伤程度,蛋白质免疫印迹法检测心肌组织中甲基转移酶样蛋白3(methyltransferase-like protein 3,METTL3)、烷基化修复同系物5(alkylation repair homolog 5,ALKBH5)及YTH结构域家族蛋白1(YTH domain family protein 1,YTHDF1)表达水平。结果与预实验假手术组比较,预实验模型组ALKBH5的表达显著下降,METTL3、YTHDF1的表达显著上调,差异有统计学意义(P<0.05)。与假手术组比较,MIRI组心肌损伤程度显著升高,miR-6321表达显著降低,METTL3和YTHDF1表达显著升高(0.96±0.06 vs 0.52±0.08,1.04±0.04 vs 0.47±0.04,P<0.05),ALKBH5表达显著降低(0.36±0.14 vs 0.89±0.09,P<0.05);与MIRI组比较,AAV-miR-6321组心肌损伤程度显著降低,miR-6321表达显著升高,METTL3和YTHDF1表达显著降低(0.58±0.05 vs 0.96±0.06,0.55±0.04 vs 1.04±0.04,P<0.05),ALKBH5表达显著升高(0.76±0.14 vs 0.36±0.14,P<0.05)。结论miR-6321可调控N6-甲基腺苷甲基化修饰从而减轻大鼠心肌缺血再灌注损伤。展开更多
BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact mo...BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.展开更多
文摘目的探讨微小RNA-6321(microRNA-6321,miR-6321)对大鼠心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)的作用,并进一步分析其作用机制与N6-甲基腺苷甲基化之间的关系。方法随机从36只雄性SD大鼠中取12只,分为预实验假手术组和预实验模型组,每组6只,用于验证MIRI模型。预实验假手术组在左前降支下方穿线而不结扎,预实验模型组结扎左前降支30 min后解开结扎线再灌注2 h。随后,本研究将剩余24只大鼠随机分为4组:假手术组、模型组(MIRI组)、腺相关病毒阴性对照组(adeno-associated virus negative control group,AAV-NC组)以及miR-6321过表达干预组(AAV-miR-6321组),每组6只。AAV-NC组和AAV-miR-6321组在模型建立前两周经尾静脉单次注射AAV-NC和AAV-miR-6321。采用冠状动脉左前降支结扎法建立大鼠MIRI模型。假手术组在左前降支下方穿线而不结扎,MIRI组、AAV-NC组和AAV-miR-6321组均进行缺血30 min、再灌注120 min处理。采用苏木精-伊红染色检测分析心肌组织损伤程度,蛋白质免疫印迹法检测心肌组织中甲基转移酶样蛋白3(methyltransferase-like protein 3,METTL3)、烷基化修复同系物5(alkylation repair homolog 5,ALKBH5)及YTH结构域家族蛋白1(YTH domain family protein 1,YTHDF1)表达水平。结果与预实验假手术组比较,预实验模型组ALKBH5的表达显著下降,METTL3、YTHDF1的表达显著上调,差异有统计学意义(P<0.05)。与假手术组比较,MIRI组心肌损伤程度显著升高,miR-6321表达显著降低,METTL3和YTHDF1表达显著升高(0.96±0.06 vs 0.52±0.08,1.04±0.04 vs 0.47±0.04,P<0.05),ALKBH5表达显著降低(0.36±0.14 vs 0.89±0.09,P<0.05);与MIRI组比较,AAV-miR-6321组心肌损伤程度显著降低,miR-6321表达显著升高,METTL3和YTHDF1表达显著降低(0.58±0.05 vs 0.96±0.06,0.55±0.04 vs 1.04±0.04,P<0.05),ALKBH5表达显著升高(0.76±0.14 vs 0.36±0.14,P<0.05)。结论miR-6321可调控N6-甲基腺苷甲基化修饰从而减轻大鼠心肌缺血再灌注损伤。
基金Supported by the Haihe Laboratory of Cell Ecosystem Innovation Fund,No.22HHXBJC00001the Key Discipline Special Project of Tianjin Municipal Health Commission,No.TJWJ2022XK016.
文摘BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.
