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
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
Objective To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro.The specific mechanism by which microRNA-144-3p is involved in Cu^(2+)-induced damage to the...Objective To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro.The specific mechanism by which microRNA-144-3p is involved in Cu^(2+)-induced damage to the human endometrial epithelial cells(HEECs)was explored.Methods HEECs were cultured in copper-containing culture medium to simulate changes in the endometrium after copper intrauterine device(Cu-IUD)implantation.Reverse transcription quantitative PCR(RT-qPCR)was used to detect the differential expression of miR-144-3p in HEECs after Cu^(2+)treatment.MiRNAs,siRNAs and related inhibitors were used to treat HEECs.The expression levels of related downstream genes were then analyzed by RT-qPCR,Western blotting and immunofluorescence to explore the specific mechanism involved.Results MiR-144-3p was significantly upregulated in the Cu^(2+)-treated HEECs.The expression of P-NF-κB,MMP9,TGF-β3 and P-SMAD3 was significantly decreased in HEECs treated with 10μg/mL Cu^(2+).MiR-144-3p regulated the expression of metallothionein 1A(MT1A)and thrombospondin-1(THBS-1)in Cu^(2+)-treated HEECs.The expression of P-NF-κB can be regulated by MT1A,and an inhibitor of P-NF-κB can significantly reduce the expression of MMP9 in Cu^(2+)-treated HEECs.The expression of TGF-β3 can be regulated by THBS-1,and a TGF-β3 inhibitor can significantly reduce the expression of SMAD3 in Cu^(2+)-treated HEECs.The proliferative capacity of HEECs treated with MMP9 or SMAD3 inhibitors was significantly reduced.Conclusions The increased Cu^(2+)concentration led to the upregulation of miR-144-3p,further reducing the expression levels of its target genes(MT1A and THBS-1),which in turn downregulated the expression of NF-κB,MMP9,TGF-β3 and SMAD3,ultimately leading to increased endometrial cell damage and decreased cell proliferation.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 探讨微小RNA(miR)-152和miR-22-3p在妊娠期糖尿病(GDM)患者外周血中的表达及意义。方法 选取2018年3月—2023年3月杭州市第九人民医院收治的96例GDM患者作为实验组,另选取健康孕妇96例作为对照组。采用qRT-PCR法检测两组孕妇血清miR-152和miR-22-3p水平及孕妇临床指标并进行比较,采用pearson法分析血清miR-152和miR-22-3p水平与临床指标相关性;Target Scan Human网站分别预测miR-152、miR-22-3p与细胞因子信号传导抑制因子3(SOCS3)的靶向关系,logistic回归分析孕妇发生GDM的影响因素;ROC曲线分析血清miR-152和miR-22-3p对孕妇是否发生GDM的预测价值。结果 GDM患者血清miR-152(1.03±0.25)和miR-22-3p(0.81±0.24)表达显著低于健康孕妇的(1.25±0.27)和(1.06±0.28),差异有统计学意义(P<0.05)。GDM患者FPG、2 h PG、FINS、TG、HbA1c、LDL-C、SOCS3、VCAM-1和HOMA-IR水平高于健康孕妇,HDL-C水平明显低于健康孕妇,差异均有统计学意义(P<0.05)。pearson分析结果表明,血清miR-152和miR-22-3p水平呈正相关(r=0.318,P<0.01),血清miR-152和miR-22-3p水平与FPG、2 h PG、FINS、TG、HbA1c、LDL-C、SOCS3、VCAM-1和HOMA-IR水平呈负相关(P<0.05)。Target Scan Human网站预测结果表明,miR-152和miR-22-3p均与SOCS3存在靶向结合位点;logistic分析结果显示miR-152、miR-22-3p、FPG、2 h PG、SOCS3、FINS、HbA1c、TG和VCAM-1均为孕妇发生GDM的影响因素(P<0.05)。ROC曲线结果表明,miR-152和miR-22-3p联合评估孕妇是否发生GDM的AUC(0.818)优于单一指标(P<0.05)。结论 GDM患者血清miR-152和miR-22-3p水平较低,两者均为孕妇发生GDM的影响因素。
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.
文摘Objective To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro.The specific mechanism by which microRNA-144-3p is involved in Cu^(2+)-induced damage to the human endometrial epithelial cells(HEECs)was explored.Methods HEECs were cultured in copper-containing culture medium to simulate changes in the endometrium after copper intrauterine device(Cu-IUD)implantation.Reverse transcription quantitative PCR(RT-qPCR)was used to detect the differential expression of miR-144-3p in HEECs after Cu^(2+)treatment.MiRNAs,siRNAs and related inhibitors were used to treat HEECs.The expression levels of related downstream genes were then analyzed by RT-qPCR,Western blotting and immunofluorescence to explore the specific mechanism involved.Results MiR-144-3p was significantly upregulated in the Cu^(2+)-treated HEECs.The expression of P-NF-κB,MMP9,TGF-β3 and P-SMAD3 was significantly decreased in HEECs treated with 10μg/mL Cu^(2+).MiR-144-3p regulated the expression of metallothionein 1A(MT1A)and thrombospondin-1(THBS-1)in Cu^(2+)-treated HEECs.The expression of P-NF-κB can be regulated by MT1A,and an inhibitor of P-NF-κB can significantly reduce the expression of MMP9 in Cu^(2+)-treated HEECs.The expression of TGF-β3 can be regulated by THBS-1,and a TGF-β3 inhibitor can significantly reduce the expression of SMAD3 in Cu^(2+)-treated HEECs.The proliferative capacity of HEECs treated with MMP9 or SMAD3 inhibitors was significantly reduced.Conclusions The increased Cu^(2+)concentration led to the upregulation of miR-144-3p,further reducing the expression levels of its target genes(MT1A and THBS-1),which in turn downregulated the expression of NF-κB,MMP9,TGF-β3 and SMAD3,ultimately leading to increased endometrial cell damage and decreased cell proliferation.
