Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).Howev...Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).However,its detailed molecular mechanism has not been adequately demonstrated.In this research,it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft(PDX)model.Mechanistically,employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis(MST),microRNA-145-5p(miR-145-5p)was pinpointed as a critical target through which elemene exerts its anti-tumor effects.Interestingly,elemene serves as a binding stabilizer for miR-145-5p,demonstrating a strong binding affinity(dissociation constant(KD)=0.39±0.17μg/mL)and preventing its degradation both in vitro and in vivo,while not interfering with the synthesis of the primary microRNA transcripts(pri-miRNAs)and precursor miRNAs(pre-miRNAs).The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA,subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated protein kinase kinase kinase 3(MAP3K3)/nuclear factor kappaB(NF-κB)pathway.Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.展开更多
目的:探讨microRNA(miR)-145是否通过调控c-Myc参与肝癌行为的调控.方法:实时定量PCR检测肝组织及细胞系miR-145表达水平;检测不同肝组织c-Myc的mRNA和蛋白表达量;通过转染上调miR-145在HepG2细胞表达后,检测转染前后空白组、实验组、...目的:探讨microRNA(miR)-145是否通过调控c-Myc参与肝癌行为的调控.方法:实时定量PCR检测肝组织及细胞系miR-145表达水平;检测不同肝组织c-Myc的mRNA和蛋白表达量;通过转染上调miR-145在HepG2细胞表达后,检测转染前后空白组、实验组、阴性对照组HepG2中c-Myc的mRNA和蛋白表达量;应用流式细胞仪检测转染前后各组HepG2的凋亡情况.结果:miR-145mRNA在正常肝组织中表达明显高于癌旁和肝癌组织(0.878±0.146vs0.265±0.084,0.271±0.096,均P<0.05);癌旁和肝癌组织中miR-145mRNA的表达量差别无统计学意义.miR-145mRNA在LO-2中的表达明显高于HepG2(0.755±0.185vs0.471±0.074,P<0.05).c-Myc mRNA和蛋白在癌旁和肝癌中表达均高于正常肝组织(mRNA:0.136±0.071,0.451±0.026vs0.029±0.023;蛋白:0.301±0.022,0.445±0.018vs0.137±0.011,均P<0.05),且在肝癌组织中表达量较癌旁组织中明显升高(P<0.05).转染miR-145mim-i c s后空白组、实验组、阴性对照组c-M y cmRNA表达差异无统计学意义;而相对于空白组和阴性对照组,实验组c-Myc蛋白表达量表达明显下降(0.146±0.011vs0.366±0.014,0.350±0.013,均P<0.05),空白组和阴性对照组之间差异无统计学意义.流式细胞术检测发现,在HepG2中上调miR-145的表达后,细胞的凋亡明显增多,实验组与空白组和阴性对照组之间有明显差异(3.000±0.100vs1.167±0.153,0.933±0.208,均P<0.05).结论:miR-145反向调节c-Myc的表达;表达下调的miR-145丧失对c-Myc的抑制可能是肝癌发生的重要机制;miR-145能够作为抑癌基因促进细胞凋亡.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.:82225048)the Dalian Science and Technology Leading Talents Project,China(Grant No.:2019RD15)Sanming Project of Medicine in Shenzhen,China(Grant No.:SZZYSM202106004).
文摘Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).However,its detailed molecular mechanism has not been adequately demonstrated.In this research,it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft(PDX)model.Mechanistically,employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis(MST),microRNA-145-5p(miR-145-5p)was pinpointed as a critical target through which elemene exerts its anti-tumor effects.Interestingly,elemene serves as a binding stabilizer for miR-145-5p,demonstrating a strong binding affinity(dissociation constant(KD)=0.39±0.17μg/mL)and preventing its degradation both in vitro and in vivo,while not interfering with the synthesis of the primary microRNA transcripts(pri-miRNAs)and precursor miRNAs(pre-miRNAs).The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA,subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated protein kinase kinase kinase 3(MAP3K3)/nuclear factor kappaB(NF-κB)pathway.Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.
文摘目的:探讨microRNA(miR)-145是否通过调控c-Myc参与肝癌行为的调控.方法:实时定量PCR检测肝组织及细胞系miR-145表达水平;检测不同肝组织c-Myc的mRNA和蛋白表达量;通过转染上调miR-145在HepG2细胞表达后,检测转染前后空白组、实验组、阴性对照组HepG2中c-Myc的mRNA和蛋白表达量;应用流式细胞仪检测转染前后各组HepG2的凋亡情况.结果:miR-145mRNA在正常肝组织中表达明显高于癌旁和肝癌组织(0.878±0.146vs0.265±0.084,0.271±0.096,均P<0.05);癌旁和肝癌组织中miR-145mRNA的表达量差别无统计学意义.miR-145mRNA在LO-2中的表达明显高于HepG2(0.755±0.185vs0.471±0.074,P<0.05).c-Myc mRNA和蛋白在癌旁和肝癌中表达均高于正常肝组织(mRNA:0.136±0.071,0.451±0.026vs0.029±0.023;蛋白:0.301±0.022,0.445±0.018vs0.137±0.011,均P<0.05),且在肝癌组织中表达量较癌旁组织中明显升高(P<0.05).转染miR-145mim-i c s后空白组、实验组、阴性对照组c-M y cmRNA表达差异无统计学意义;而相对于空白组和阴性对照组,实验组c-Myc蛋白表达量表达明显下降(0.146±0.011vs0.366±0.014,0.350±0.013,均P<0.05),空白组和阴性对照组之间差异无统计学意义.流式细胞术检测发现,在HepG2中上调miR-145的表达后,细胞的凋亡明显增多,实验组与空白组和阴性对照组之间有明显差异(3.000±0.100vs1.167±0.153,0.933±0.208,均P<0.05).结论:miR-145反向调节c-Myc的表达;表达下调的miR-145丧失对c-Myc的抑制可能是肝癌发生的重要机制;miR-145能够作为抑癌基因促进细胞凋亡.
