Despite androgen dependence in a majority of castration-resistant prostate cancers,some cancer cells are independent of androgen receptor(AR)function,a feature of heterogeneity in prostate cancer.One of the aggressive...Despite androgen dependence in a majority of castration-resistant prostate cancers,some cancer cells are independent of androgen receptor(AR)function,a feature of heterogeneity in prostate cancer.One of the aggressive variants of prostate cancer that are AR independent is neuroendocrine prostate cancer(NEPC).This manuscript will focus on the new finding of human one cut domain family member 2(ONECUT2)transcription factor and its role in castration resistance,especially in NEPC.展开更多
目的:探讨miR-9-5p在乳腺癌恶性生物学行为中发挥的作用及其可能的调控机制。方法:利用OncomiR在线数据库分析miR-9-5p在乳腺癌组织与正常乳腺组织中表达的差异,qPCR检测乳腺癌细胞系与正常乳腺细胞中miR-9-5p表达水平。基于靶基因预测...目的:探讨miR-9-5p在乳腺癌恶性生物学行为中发挥的作用及其可能的调控机制。方法:利用OncomiR在线数据库分析miR-9-5p在乳腺癌组织与正常乳腺组织中表达的差异,qPCR检测乳腺癌细胞系与正常乳腺细胞中miR-9-5p表达水平。基于靶基因预测软件TargetScan分析ONECUT2(one cut homeobox 2)可能是miR-9-5p的作用靶基因,双荧光素酶报告实验验证两者的靶向关系。向MDA-231细胞中分别转染miR-9-5p mimic、ONECUT2 siRNA及相应对照,qPCR及WB实验检测转染对MDA-231细胞中干性基因NOTCH1、NANOG和Y染色体性别决定区(sex-determing region of Y chromosome,SRY)-盒转录因子9(SRY-box transcription factor 9,SOX9)表达水平的影响,BrdU法、AnnexinⅤ流式细胞术、MTS实验分别检测转染对细胞增殖、凋亡和化疗耐药的影响,ALDEFLUOR染色流式细胞术检测miR-9-5p及靶基因ONECUT2对肿瘤干细胞化特征的影响。建立NSG小鼠乳腺癌化疗模型,体内实验进一步验证ONECUT2对肿瘤干性化及化疗抵抗等肿瘤恶性生物学行为的影响。结果:miR-9-5p在乳腺癌组织(P=0.007)及乳腺癌MDA-231细胞系(P=0.0005)中呈现显著高表达,并与乳腺癌患者不良预后呈正相关(P=0.0016)。miR-9-5p可靶向负调控ONECUT2,进而增加ALDH+MDA-231细胞比例(P=0.0006),上调干性NOTCH1、NANOG和SOX9蛋白表达,并增强乳腺癌细胞抗凋亡能力(P=0.0003)及其对多西他赛(DTX)和多柔比星(DOXO)化疗的耐受性;然而miR-9-5p/ONECUT2轴未能显著影响MDA-231细胞的增殖能力(P>0.05)。与对照组相比,MDA-231/ONECUT2组小鼠接受DTX治疗后,移植瘤体积较对照组显著缩小(P<0.05),瘤组织中NOTCH1、SOX9蛋白和ABC转运蛋白的mRNA和蛋白表达水平均显著降低(P<0.05或P<0.01)。结论:乳腺癌组织中高表达的miR-9-5p通过靶向ONECUT2诱导乳腺癌干细胞化及抗凋亡能力,增强了其对化学治疗的抵抗性。展开更多
文摘Despite androgen dependence in a majority of castration-resistant prostate cancers,some cancer cells are independent of androgen receptor(AR)function,a feature of heterogeneity in prostate cancer.One of the aggressive variants of prostate cancer that are AR independent is neuroendocrine prostate cancer(NEPC).This manuscript will focus on the new finding of human one cut domain family member 2(ONECUT2)transcription factor and its role in castration resistance,especially in NEPC.
文摘目的:探讨miR-9-5p在乳腺癌恶性生物学行为中发挥的作用及其可能的调控机制。方法:利用OncomiR在线数据库分析miR-9-5p在乳腺癌组织与正常乳腺组织中表达的差异,qPCR检测乳腺癌细胞系与正常乳腺细胞中miR-9-5p表达水平。基于靶基因预测软件TargetScan分析ONECUT2(one cut homeobox 2)可能是miR-9-5p的作用靶基因,双荧光素酶报告实验验证两者的靶向关系。向MDA-231细胞中分别转染miR-9-5p mimic、ONECUT2 siRNA及相应对照,qPCR及WB实验检测转染对MDA-231细胞中干性基因NOTCH1、NANOG和Y染色体性别决定区(sex-determing region of Y chromosome,SRY)-盒转录因子9(SRY-box transcription factor 9,SOX9)表达水平的影响,BrdU法、AnnexinⅤ流式细胞术、MTS实验分别检测转染对细胞增殖、凋亡和化疗耐药的影响,ALDEFLUOR染色流式细胞术检测miR-9-5p及靶基因ONECUT2对肿瘤干细胞化特征的影响。建立NSG小鼠乳腺癌化疗模型,体内实验进一步验证ONECUT2对肿瘤干性化及化疗抵抗等肿瘤恶性生物学行为的影响。结果:miR-9-5p在乳腺癌组织(P=0.007)及乳腺癌MDA-231细胞系(P=0.0005)中呈现显著高表达,并与乳腺癌患者不良预后呈正相关(P=0.0016)。miR-9-5p可靶向负调控ONECUT2,进而增加ALDH+MDA-231细胞比例(P=0.0006),上调干性NOTCH1、NANOG和SOX9蛋白表达,并增强乳腺癌细胞抗凋亡能力(P=0.0003)及其对多西他赛(DTX)和多柔比星(DOXO)化疗的耐受性;然而miR-9-5p/ONECUT2轴未能显著影响MDA-231细胞的增殖能力(P>0.05)。与对照组相比,MDA-231/ONECUT2组小鼠接受DTX治疗后,移植瘤体积较对照组显著缩小(P<0.05),瘤组织中NOTCH1、SOX9蛋白和ABC转运蛋白的mRNA和蛋白表达水平均显著降低(P<0.05或P<0.01)。结论:乳腺癌组织中高表达的miR-9-5p通过靶向ONECUT2诱导乳腺癌干细胞化及抗凋亡能力,增强了其对化学治疗的抵抗性。
