探讨miRNA-21-3p在急性髓系白血病(AML)患者外周血单个核细胞(PBMC)中的表达水平变化,并分析其临床意义。方法:采用qRT-PCR法测定正常对照和AML患者PBMC的miRNA-21-3p表达水平,根据ROC曲线计算miRNA-21-3p临界值(cut-off值)。体外CCK8...探讨miRNA-21-3p在急性髓系白血病(AML)患者外周血单个核细胞(PBMC)中的表达水平变化,并分析其临床意义。方法:采用qRT-PCR法测定正常对照和AML患者PBMC的miRNA-21-3p表达水平,根据ROC曲线计算miRNA-21-3p临界值(cut-off值)。体外CCK8细胞计数法测定白血病细胞的增殖水平。分别比较不同患者组的miRNA-21-3p表达水平,并根据cut-off值分为大于和小于两组,同时分析其临床预后意义。结果:①急性髓系白血病(AML)患者与正常对照组比较,PBMCmiRNA-21-3p平均表达水平高于正常对照组,其临界cut-of值为1.426^(2^-△△Ct)。②不同组白血病患者miRNA-21-3p水平的比较:未复发组AML患者的PBMC O miRNA-21-3p水平明显低于复发组(P=0.0027),完全缓解(CR)组与未缓解(NR)组的PBMCmiR-21-3p水平差别不明显(t=1.254,P=0.2146),MRD(+)组PBMCmiRNA-21-3p水平高于MRD(-)组(P=0.0001)。③大于和小于临界值两组的临床意义分析:PBMCmiRNA-21-3p小于临界值组的无病生存期(DFS)与大于临界值组无明显差异(P=0.2331),大于临界值组的复发率和微小残留病(MRD)阳性率均高于小于临界值组(P<0.0001)。大于临界值组和低于临界值组相比较,完全缓解率无明显差异,但是PBMCmiRNA-21-3p大于临界值组的5年生存率低于小于临界值组(P=0.0109)。④CCK8细胞计数法测定miRNA-21-3pmimics对白血病细胞增殖的影响。结果表明miR-21-3pmimics对U937、THP1和K562细胞的增殖水平并没有明显的直接作用(P>0.05)。结论:PBMCmiRNA-21-3p水平与白血病预后有关,并以大于和小于临界值分组更为敏感。miRNA-21-3p mimics对U937、THP1和K562细胞的增殖水平没有明显的直接作用。展开更多
目的探究血清microRNA-21(miR-21)、microRNA-193a-3p(miR-193a-3p)水平与结直肠癌患者手术预后的关系。方法回顾性分析2020年1月—2022年1月苏州大学附属第一医院收治112例结直肠癌患者的病历资料。患者均接受结直肠癌根治术,术后随访1...目的探究血清microRNA-21(miR-21)、microRNA-193a-3p(miR-193a-3p)水平与结直肠癌患者手术预后的关系。方法回顾性分析2020年1月—2022年1月苏州大学附属第一医院收治112例结直肠癌患者的病历资料。患者均接受结直肠癌根治术,术后随访16个月,记录患者的预后生存结局,多因素逐步Logistic回归分析结直肠癌患者手术预后的影响因素,评估血清miR-21、miR-193a-3p对结直肠癌患者预后的预测效能。结果112例结直肠癌患者死亡22例,病死率为19.64%;生存90例,生存率为80.36%。死亡组术前血清miR-21 mRNA相对表达量、临床分期Ⅲ期占比、淋巴结转移率均高于生存组(P<0.05),血清miR-193a-3p m RNA相对表达量低于生存组(P<0.05)。多因素逐步Logistic回归分析结果显示,临床分期Ⅲ期[OR=3.777(95%CI:1.399,10.194)]、淋巴结转移[OR=5.099(95%CI:1.715,15.156)]、miR-21表达升高[OR=4.889(95%CI:1.645,14.533)]、miR-193a-3p表达降低[OR=4.402(95%CI:1.481,13.084)]均是直肠癌患者预后的影响因素(P<0.05)。受试者工作特性曲线分析结果显示,血清miR-21、miR-193a-3p单一及联合预测结直肠癌预后的敏感性分别为69.04%(95%CI:0.487,0.813)、72.73%(95%CI:0.495,0.884)、86.36%(95%CI:0.640,0.964),特异性分别为62.22%(95%CI:0.513,0.720)、68.89%(95%CI:0.581,0.780)、90.00%(95%CI:0.814,0.950),曲线下面积分别为0.782、0.731和0.901。结论结直肠癌患者术前miR-21、miR-193a-3p表达与术后预后密切相关,且在结直肠癌患者的预后结局中表现出良好的预测效能。展开更多
目的探讨茵陈蒿汤对代谢相关脂肪性肝病(MAFLD)小鼠LncMEG3/miR-21-5p/PPARα介导脂肪酸β氧化的影响。方法为探讨MAFLD差异表达的miRNAs,应用PubMed、Web of Science和中国知网3个电子数据库对以中英文发表的研究进行检索,采用随机效...目的探讨茵陈蒿汤对代谢相关脂肪性肝病(MAFLD)小鼠LncMEG3/miR-21-5p/PPARα介导脂肪酸β氧化的影响。方法为探讨MAFLD差异表达的miRNAs,应用PubMed、Web of Science和中国知网3个电子数据库对以中英文发表的研究进行检索,采用随机效应模型进行Meta分析。24只C57BL/6J小鼠,SPF级,采用随机数字表法将小鼠随机分为对照组、模型组和茵陈蒿汤组,共3组,每组8只。采用喂饲高脂饲料的方法对模型组和茵陈蒿汤组小鼠建立MAFLD模型。12周后,茵陈蒿汤组给予茵陈蒿汤灌胃,每日1次,连续4周。采用HE染色法和油红O染色法观察各组小鼠肝组织病理形态表现;全自动生化分析仪检测血清甘油三酯(TG)、总胆固醇(TC)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、高密度脂蛋白胆固(HDL-C)和低密度脂蛋白胆固醇(LDL-C);试剂盒检测小鼠肝脏甘油三酯(TG)和游离脂肪酸(FFA)水平;RT-qPCR法检测小鼠LncMEG3、miR-21-5p、miR-34a-5p、miR-122-5p、miR-192-5p、过氧化物酶体增殖物激活受体α(PPARα)、肉碱棕榈酰转移酶1A(CPT1A)、肉碱棕榈酰基转移酶2(CPT2)、中链酰基辅酶A脱氢酶(MCAD)、长链脂酰辅酶A脱氢酶(LCAD)、酰基辅酶A氧化酶1(ACOX1)和三羟酰辅酶A脱氢酶(EHHADH)mRNA水平蛋白表达水平。结果Meta分析结果显示,与健康对照组相比,MAFLD组miR-34a-5p、miR-122-5p、miR-192-5p、miR-21-5p、miR-451-5p水平显著升高。肝脏HE染色结果发现,模型组小鼠出现肝细胞肿胀,体积增大,有明显脂肪变性;与模型组相比,茵陈蒿汤组小鼠肝细胞体积肿胀明显降低,脂肪变性显著减少。肝脏油红O染色结果发现,与对照组相比,模型组肝脏可见大量橘红色脂滴,脂质沉积显著升高;茵陈蒿汤组可见少量脂肪变性,橘红色脂滴分布减少,肝脏脂质沉积情况明显改善。相较于对照组,模型组小鼠血清HDL-C、LDL-C、TG、TC、AST、ALT表达显著升高(P<0.05),肝脏TG和FFA水平均显著升高(P<0.05);相较于模型组,茵陈蒿汤组小鼠肝脏FFA和TG水平显著降低(P<0.05),血清AST、LDL-C、TG、ALT水平显著降低(P<0.05)。与对照组相比,模型组miR-21-5p、miR-34a-5p、miR-122-5p、miR-192-5p表达水平增高(P<0.05);与模型组相比,茵陈蒿汤组miR-21-5p、miR-34a-5p、miR-122-5p、miR-192-5p表达水平降低(P<0.05),miR-21-5p变化倍数(模型组/茵陈蒿汤组)最大。与对照组相比,模型组Lnc MEG3表达水平降低(P<0.05);与模型组相比,茵陈蒿汤组Lnc MEG3表达水平升高(P<0.05)。与对照组相比,模型组PPARα、CPT1A、CPT2、MCAD、LCAD、ACOX1、EHHADH mRNA和蛋白表达水平显著降低(P<0.05);与模型组相比,茵陈蒿汤组PPARα、CPT1A、CPT2、MCAD、LCAD、ACOX1、EHHADH mRNA和蛋白表达水平显著升高(P<0.05)。结论茵陈蒿汤可能通过调控Lnc MEG3/miR-21-5p/PPARα信号通路促进肝脏脂肪酸β氧化,从而达到防治MAFLD的作用。展开更多
AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)...AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.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
文摘探讨miRNA-21-3p在急性髓系白血病(AML)患者外周血单个核细胞(PBMC)中的表达水平变化,并分析其临床意义。方法:采用qRT-PCR法测定正常对照和AML患者PBMC的miRNA-21-3p表达水平,根据ROC曲线计算miRNA-21-3p临界值(cut-off值)。体外CCK8细胞计数法测定白血病细胞的增殖水平。分别比较不同患者组的miRNA-21-3p表达水平,并根据cut-off值分为大于和小于两组,同时分析其临床预后意义。结果:①急性髓系白血病(AML)患者与正常对照组比较,PBMCmiRNA-21-3p平均表达水平高于正常对照组,其临界cut-of值为1.426^(2^-△△Ct)。②不同组白血病患者miRNA-21-3p水平的比较:未复发组AML患者的PBMC O miRNA-21-3p水平明显低于复发组(P=0.0027),完全缓解(CR)组与未缓解(NR)组的PBMCmiR-21-3p水平差别不明显(t=1.254,P=0.2146),MRD(+)组PBMCmiRNA-21-3p水平高于MRD(-)组(P=0.0001)。③大于和小于临界值两组的临床意义分析:PBMCmiRNA-21-3p小于临界值组的无病生存期(DFS)与大于临界值组无明显差异(P=0.2331),大于临界值组的复发率和微小残留病(MRD)阳性率均高于小于临界值组(P<0.0001)。大于临界值组和低于临界值组相比较,完全缓解率无明显差异,但是PBMCmiRNA-21-3p大于临界值组的5年生存率低于小于临界值组(P=0.0109)。④CCK8细胞计数法测定miRNA-21-3pmimics对白血病细胞增殖的影响。结果表明miR-21-3pmimics对U937、THP1和K562细胞的增殖水平并没有明显的直接作用(P>0.05)。结论:PBMCmiRNA-21-3p水平与白血病预后有关,并以大于和小于临界值分组更为敏感。miRNA-21-3p mimics对U937、THP1和K562细胞的增殖水平没有明显的直接作用。
文摘目的探究血清microRNA-21(miR-21)、microRNA-193a-3p(miR-193a-3p)水平与结直肠癌患者手术预后的关系。方法回顾性分析2020年1月—2022年1月苏州大学附属第一医院收治112例结直肠癌患者的病历资料。患者均接受结直肠癌根治术,术后随访16个月,记录患者的预后生存结局,多因素逐步Logistic回归分析结直肠癌患者手术预后的影响因素,评估血清miR-21、miR-193a-3p对结直肠癌患者预后的预测效能。结果112例结直肠癌患者死亡22例,病死率为19.64%;生存90例,生存率为80.36%。死亡组术前血清miR-21 mRNA相对表达量、临床分期Ⅲ期占比、淋巴结转移率均高于生存组(P<0.05),血清miR-193a-3p m RNA相对表达量低于生存组(P<0.05)。多因素逐步Logistic回归分析结果显示,临床分期Ⅲ期[OR=3.777(95%CI:1.399,10.194)]、淋巴结转移[OR=5.099(95%CI:1.715,15.156)]、miR-21表达升高[OR=4.889(95%CI:1.645,14.533)]、miR-193a-3p表达降低[OR=4.402(95%CI:1.481,13.084)]均是直肠癌患者预后的影响因素(P<0.05)。受试者工作特性曲线分析结果显示,血清miR-21、miR-193a-3p单一及联合预测结直肠癌预后的敏感性分别为69.04%(95%CI:0.487,0.813)、72.73%(95%CI:0.495,0.884)、86.36%(95%CI:0.640,0.964),特异性分别为62.22%(95%CI:0.513,0.720)、68.89%(95%CI:0.581,0.780)、90.00%(95%CI:0.814,0.950),曲线下面积分别为0.782、0.731和0.901。结论结直肠癌患者术前miR-21、miR-193a-3p表达与术后预后密切相关,且在结直肠癌患者的预后结局中表现出良好的预测效能。
文摘AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.
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.
