BACKGROUND Multinucleated giant cells(MGCs)in bladder carcinomas are poorly studied.AIM To describe the function,morphogenesis,and origin of mononuclear and MGCs in urothelial carcinoma(UC)of the bladder in Bulgarian ...BACKGROUND Multinucleated giant cells(MGCs)in bladder carcinomas are poorly studied.AIM To describe the function,morphogenesis,and origin of mononuclear and MGCs in urothelial carcinoma(UC)of the bladder in Bulgarian and French patients.METHODS Urothelial bladder carcinomas(n=104)from 2016-2020 were analyzed retrospectively using immunohistochemical(IHC)and histochemical stain examination.Giant cells in the bladder stroma were found in 35.6%of cases,more often in highgrades.RESULTS We confirm that MGCs in the mucosa in UC of the bladder were positive for both mesenchymal and myofibroblast markers(vimentin,smooth muscle actin,Desmin,and CD34)and the macrophage marker CD68.Furthermore,IHC studies revealed the following profile of these cells:Positive for p16;negative for epithelial(CK AE1/AE3 and GATA-3),vascular(CD31),neural(PS100 and CKIT),cambial,blastic(CD34-blasts and C-KIT),and immune markers(IG G,immunoglobulin G4,and PD-L1);no proliferative activity,possess no specific immune function,and cannot be used to calculate the Combined Positive Score scale.CONCLUSION In conclusion,the giant stromal cells in non-tumor and tumor bladder can be used as a characteristic and relatively constant,although nonspecific,histological marker for chronic bladder damage,reflecting the chronic irritation or inflammation.Likewise,according to the morphological and IHC of the mono-and multinucleated giant cells in the bladder,they are most likely represent telocytes capable of adapting their morphology to the pathology of the organ.展开更多
目的探讨骨髓间充质干细胞(BMSCs)在平滑肌微环境中发生分化后平滑肌标志性基因乙酰化水平的变化,以及组蛋白乙酰化修饰在干细胞分化中的作用及机制。方法体外培养BMSCs和膀胱平滑肌细胞(BSMCs),选择同批次的第3代BMSCs,将与BSMCs共培...目的探讨骨髓间充质干细胞(BMSCs)在平滑肌微环境中发生分化后平滑肌标志性基因乙酰化水平的变化,以及组蛋白乙酰化修饰在干细胞分化中的作用及机制。方法体外培养BMSCs和膀胱平滑肌细胞(BSMCs),选择同批次的第3代BMSCs,将与BSMCs共培养3d的BMSCs作为实验组,未经共培养的BMSCs作为对照组,采用RT-PCR检测两组BMSCs中平滑肌α肌动蛋白(α-SMA)、钙调节蛋白(calponin)、平滑肌肌球蛋白重链(SM-MHC)的表达丰度。采用条件为80%、20次、0.5 s、8个循环的超声破碎实验组及对照组BMSCs的DNA,以H3K9抗体结合特定乙酰化位点,采用免疫共沉淀技术(ChIP)沉淀BMSCs组蛋白乙酰化位点基因,接头PCR扩增所获基因,Real-time PCR检测所获基因中3种目的基因(α-SMA、Calponin、SM-MHC)的表达水平。结果 RT-PCR检测结果显示,实验组BMSCs中的平滑肌标志性基因(α-SMA、Calponin、SM-MHC)的mRNA表达水平较对照组明显增高[分别为0.176±0.003 vs. 0.070±0.002,0.079±0.002vs.0.051±0.003,0.091±0.004vs.0.034±0.001],差异均有统计学意义(P<0.01)。分光光度计检测ChIP后获得的DNA,结果表明H3K9乙酰化抗体沉淀所获DNA浓度高于Ig G抗体,且实验组高于对照组(P<0.05)。RealtimePCR分析BMSCs分化前后目的基因组蛋白H3K9乙酰化水平,结果显示,BMSCs分化后,实验组的平滑肌标志性基因α-SMA、calponin、SM-MHC的mRNA转录水平明显高于对照组[分别为9.26±5.03 vs. 1.01±0.05,2.33±0.65 vs.0.99±0.05,2.63±0.37vs.1.00±0.03],差异均有统计学意义(P>0.05)。结论在平滑肌微环境中,BSMCs特定位点H3K9乙酰化程度的增加可促进BMSCs向BSMCs的分化。展开更多
基金the European Union-NextGenerationEU,through the National Recovery and Resilience Plan of the Republic of Bulgaria,No.BG-RRP-2.004-0008.
文摘BACKGROUND Multinucleated giant cells(MGCs)in bladder carcinomas are poorly studied.AIM To describe the function,morphogenesis,and origin of mononuclear and MGCs in urothelial carcinoma(UC)of the bladder in Bulgarian and French patients.METHODS Urothelial bladder carcinomas(n=104)from 2016-2020 were analyzed retrospectively using immunohistochemical(IHC)and histochemical stain examination.Giant cells in the bladder stroma were found in 35.6%of cases,more often in highgrades.RESULTS We confirm that MGCs in the mucosa in UC of the bladder were positive for both mesenchymal and myofibroblast markers(vimentin,smooth muscle actin,Desmin,and CD34)and the macrophage marker CD68.Furthermore,IHC studies revealed the following profile of these cells:Positive for p16;negative for epithelial(CK AE1/AE3 and GATA-3),vascular(CD31),neural(PS100 and CKIT),cambial,blastic(CD34-blasts and C-KIT),and immune markers(IG G,immunoglobulin G4,and PD-L1);no proliferative activity,possess no specific immune function,and cannot be used to calculate the Combined Positive Score scale.CONCLUSION In conclusion,the giant stromal cells in non-tumor and tumor bladder can be used as a characteristic and relatively constant,although nonspecific,histological marker for chronic bladder damage,reflecting the chronic irritation or inflammation.Likewise,according to the morphological and IHC of the mono-and multinucleated giant cells in the bladder,they are most likely represent telocytes capable of adapting their morphology to the pathology of the organ.
文摘目的探讨骨髓间充质干细胞(BMSCs)在平滑肌微环境中发生分化后平滑肌标志性基因乙酰化水平的变化,以及组蛋白乙酰化修饰在干细胞分化中的作用及机制。方法体外培养BMSCs和膀胱平滑肌细胞(BSMCs),选择同批次的第3代BMSCs,将与BSMCs共培养3d的BMSCs作为实验组,未经共培养的BMSCs作为对照组,采用RT-PCR检测两组BMSCs中平滑肌α肌动蛋白(α-SMA)、钙调节蛋白(calponin)、平滑肌肌球蛋白重链(SM-MHC)的表达丰度。采用条件为80%、20次、0.5 s、8个循环的超声破碎实验组及对照组BMSCs的DNA,以H3K9抗体结合特定乙酰化位点,采用免疫共沉淀技术(ChIP)沉淀BMSCs组蛋白乙酰化位点基因,接头PCR扩增所获基因,Real-time PCR检测所获基因中3种目的基因(α-SMA、Calponin、SM-MHC)的表达水平。结果 RT-PCR检测结果显示,实验组BMSCs中的平滑肌标志性基因(α-SMA、Calponin、SM-MHC)的mRNA表达水平较对照组明显增高[分别为0.176±0.003 vs. 0.070±0.002,0.079±0.002vs.0.051±0.003,0.091±0.004vs.0.034±0.001],差异均有统计学意义(P<0.01)。分光光度计检测ChIP后获得的DNA,结果表明H3K9乙酰化抗体沉淀所获DNA浓度高于Ig G抗体,且实验组高于对照组(P<0.05)。RealtimePCR分析BMSCs分化前后目的基因组蛋白H3K9乙酰化水平,结果显示,BMSCs分化后,实验组的平滑肌标志性基因α-SMA、calponin、SM-MHC的mRNA转录水平明显高于对照组[分别为9.26±5.03 vs. 1.01±0.05,2.33±0.65 vs.0.99±0.05,2.63±0.37vs.1.00±0.03],差异均有统计学意义(P>0.05)。结论在平滑肌微环境中,BSMCs特定位点H3K9乙酰化程度的增加可促进BMSCs向BSMCs的分化。
