目的探讨ADAM12L在基底细胞样乳腺癌组织中的蛋白和核酸表达水平,并分析其临床意义。方法应用免疫组化法检测86例基底细胞样乳腺癌组织和20例良性乳腺纤维瘤组织的ADAM12L蛋白表达水平,并进一步采用实时荧光定量PCR方法检测60例基底细...目的探讨ADAM12L在基底细胞样乳腺癌组织中的蛋白和核酸表达水平,并分析其临床意义。方法应用免疫组化法检测86例基底细胞样乳腺癌组织和20例良性乳腺纤维瘤组织的ADAM12L蛋白表达水平,并进一步采用实时荧光定量PCR方法检测60例基底细胞样乳腺癌组织及其配对癌旁组织的ADAM12L m RNA表达量。结果免疫组化染色显示,基底细胞样乳腺癌标本中ADAM12L蛋白阳性表达率(84.9%,73/86)显著高于良性纤维瘤组织样本中ADAM12L蛋白阳性表达率(45%,9/20),P<0.05;淋巴结转移阳性者的ADAM12L阳性率(73.8%,31/42)明显高于无淋巴结转移者(47.8%,21/44),χ~2=5.073,P=0.024;临床分期Ⅲ~Ⅳ者,ADAM12L阳性率(74.5%,41/55)明显高于临床分期Ⅰ~Ⅱ者(35.5%,11/31),χ~2=11.073,P<0.01。real-time PCR检测结果显示ADAM12L在基底细胞样乳腺癌组织中的表达量为2.07±0.95,显著高于配对癌旁组织的表达量(P<0.05)。结论基底细胞样乳腺癌组织的ADAM12L蛋白与核酸表达量均显著高于正常或良性乳腺组织,且与乳腺癌淋巴结转移和临床分期有着密切的联系。ADAM12L可能可以作为新的基底细胞样乳腺癌患者的个体化诊疗靶点。展开更多
Objective Pituitary adenomas(PAs)can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation.Previous studies demonstrated that excessive expression of metalloproteases AD...Objective Pituitary adenomas(PAs)can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation.Previous studies demonstrated that excessive expression of metalloproteases ADAM12 and MMP-14 is instrumental for the active proliferation and invasiveness of PA cells in vitro and of tumors in vivo.However,the mechanisms regulating ADAM12 and MMP-14 expression in PAs remain unclear.Methods Target gene prediction and transcriptomic profiling of invasive vs.noninvasive human PA samples were performed to identify miRNA species potentially involved in the regulation of ADAM12 and MMP14.For cellular analyses of miRNA functions,two mouse PA cell lines(AtT20 and TtT/GF)were transfected with miR-149-3p and miR-149-5p,respectively.The effects of miR-149(3p and 5p)on expression levels of ADAM12 and MMP14 were determined by Western blotting followed by an analysis of proliferation and colony formation assays,scratch migration assays,and invasion assays.Results A significant downregulation of miRNA-149 was observed in invasive vs.noninvasive PA(0.32 vs.0.09,P<0.0001).In AtT-20 and TtT/GF mouse PAs cells,transfection of mimic miRNA-149(3p and 5p)caused a significantly reduced cell proliferation and matrigel invasion,whilst the effect on cell migration was less pronounced.Both strands of miRNA-149(3p and 5p)markedly reduced protein levels of ADAM12 and MMP-14 by at least 40%in both cell lines.Conclusion This study proved that the invasiveness of PA cells is,at least partly,regulated by miRNA-149-dependent expression of ADAM12 and MMP-14.展开更多
文摘目的探讨ADAM12L在基底细胞样乳腺癌组织中的蛋白和核酸表达水平,并分析其临床意义。方法应用免疫组化法检测86例基底细胞样乳腺癌组织和20例良性乳腺纤维瘤组织的ADAM12L蛋白表达水平,并进一步采用实时荧光定量PCR方法检测60例基底细胞样乳腺癌组织及其配对癌旁组织的ADAM12L m RNA表达量。结果免疫组化染色显示,基底细胞样乳腺癌标本中ADAM12L蛋白阳性表达率(84.9%,73/86)显著高于良性纤维瘤组织样本中ADAM12L蛋白阳性表达率(45%,9/20),P<0.05;淋巴结转移阳性者的ADAM12L阳性率(73.8%,31/42)明显高于无淋巴结转移者(47.8%,21/44),χ~2=5.073,P=0.024;临床分期Ⅲ~Ⅳ者,ADAM12L阳性率(74.5%,41/55)明显高于临床分期Ⅰ~Ⅱ者(35.5%,11/31),χ~2=11.073,P<0.01。real-time PCR检测结果显示ADAM12L在基底细胞样乳腺癌组织中的表达量为2.07±0.95,显著高于配对癌旁组织的表达量(P<0.05)。结论基底细胞样乳腺癌组织的ADAM12L蛋白与核酸表达量均显著高于正常或良性乳腺组织,且与乳腺癌淋巴结转移和临床分期有着密切的联系。ADAM12L可能可以作为新的基底细胞样乳腺癌患者的个体化诊疗靶点。
基金This research was funded in part by the ERANET PerMed consortium“PerProGlio”(to J.W.B.and A.S.).
文摘Objective Pituitary adenomas(PAs)can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation.Previous studies demonstrated that excessive expression of metalloproteases ADAM12 and MMP-14 is instrumental for the active proliferation and invasiveness of PA cells in vitro and of tumors in vivo.However,the mechanisms regulating ADAM12 and MMP-14 expression in PAs remain unclear.Methods Target gene prediction and transcriptomic profiling of invasive vs.noninvasive human PA samples were performed to identify miRNA species potentially involved in the regulation of ADAM12 and MMP14.For cellular analyses of miRNA functions,two mouse PA cell lines(AtT20 and TtT/GF)were transfected with miR-149-3p and miR-149-5p,respectively.The effects of miR-149(3p and 5p)on expression levels of ADAM12 and MMP14 were determined by Western blotting followed by an analysis of proliferation and colony formation assays,scratch migration assays,and invasion assays.Results A significant downregulation of miRNA-149 was observed in invasive vs.noninvasive PA(0.32 vs.0.09,P<0.0001).In AtT-20 and TtT/GF mouse PAs cells,transfection of mimic miRNA-149(3p and 5p)caused a significantly reduced cell proliferation and matrigel invasion,whilst the effect on cell migration was less pronounced.Both strands of miRNA-149(3p and 5p)markedly reduced protein levels of ADAM12 and MMP-14 by at least 40%in both cell lines.Conclusion This study proved that the invasiveness of PA cells is,at least partly,regulated by miRNA-149-dependent expression of ADAM12 and MMP-14.
