Nephroblastoma is one of the most common causes of abdominal-pelvic mass in children. It still raises a diagnostic problem because of the wide variety of causes of abdominal-pelvic mass but also because of the delays ...Nephroblastoma is one of the most common causes of abdominal-pelvic mass in children. It still raises a diagnostic problem because of the wide variety of causes of abdominal-pelvic mass but also because of the delays in consultation in sub-Saharan black Africa and consequently the discovery of the mass at a very late and sometimes metastatic stage. Yet nephroblastoma is a very chemo-sensitive malignancy requiring diagnosis at an early stage;a procedure in which medical imaging is essential. We report the case of a large abdominal-pelvic mass in a three-year-old girl in whom the abdominal-pelvic CT allowed to diagnose nephroblastoma with liver metastases. Our objective is to demonstrate the contribution of computed tomography in the diagnosis of large abdominal-pelvic mass of the child and discuss other causes of abdominal-pelvic mass of the child.展开更多
The overall frequency of WT1 gene alterations in Wilms tumor is still unclear in Taiwan. Here we conducted molecular genetic analysis of the WT1 gene in Taiwan Residents patients with Wilms tumor. Polymerase chain rea...The overall frequency of WT1 gene alterations in Wilms tumor is still unclear in Taiwan. Here we conducted molecular genetic analysis of the WT1 gene in Taiwan Residents patients with Wilms tumor. Polymerase chain reaction and direct sequencing were performed on DNA samples from blood and paraffin-embedded tumor specimens. A constitutional mutation in the WT1 gene was found in one DNA sample from peripheral blood lymphocytes. The remaining DNA samples from peripheral blood lymphocytes and paraffin-embedded tumor specimens were tested negative for both constitutional mutations and somatic mutations. Thus, mutations at other Wilms tumor loci may play an important role in Wilms tumor development.展开更多
Objective To explore the effects of bufalin on inhibiting proliferation, up-regulating methylation of Wilm’ tumor 1 gene (WT1) as well as its possible mechanisms in human erythroid leukemic (HEL) cells. Methods The H...Objective To explore the effects of bufalin on inhibiting proliferation, up-regulating methylation of Wilm’ tumor 1 gene (WT1) as well as its possible mechanisms in human erythroid leukemic (HEL) cells. Methods The HEL cells were treated with bufalin at various concentrations to observe cellular morphology, proliferation assay and cell cycle. The mRNA and protein expression levels of WT1 were detected by reverse transcription polymerase chain reaction (RT-PCR), Western blot and immunocytochemistry, DNA methylation of WT1 and protein expression levels of DNA methyltransferase 3a (DNMT3a) and DNMT3b were analyzed by methylation-specific PCR, and Western blot respectively. Results The bufalin was effective to inhibit proliferation of HEL cells in a dose-dependent manner, their suppression rates were from 23.4%±2.1% to 87.2%±5.4% with an half maximal inhibit concentration (IC<sub>50</sub>) of 0.046 μmol/L. Typical apoptosis morphology was observed in bufalin-treated HEL cells. The proliferation index of cell cycle decreased from 76.4%±1.9% to 49.7%±1.3%. The expression levels of WT1 mRNA and its protein reduced gradually with increasing doses of bufalin, meanwhile, the methylation status of WT1 gene changed from unmethylated into partially or totally methylated. While, the expression levels of DNMT3a and DNMT3b protein gradually increased by bufalin treatment in a dose-dependent manner. Conclusions Bufalin can not only significantly inhibit the proliferation of HEL cells and arrest cell cycle at G<sub>0</sub>/G<sub>1</sub> phase, but also induce cellular apoptosis and down-regulate the expression level of WT1. Our results provide the evidence of bufalin for anti-leukemia, its mechanism may involve in increasing WT1 methylation status which is related to the up-regulation of DNMT3a and DNMT3b proteins in erythroid leukemic HEL cells.展开更多
肾母细胞瘤(Wilms tumor,WT)是儿童最常见的肾恶性肿瘤。COG(Children’s Oncology Group,儿童肿瘤协作组)和SIOP(International Society of Paediatric Oncology,国际儿童肿瘤学会)两大儿童肿瘤研究的国际协作组织分别形成了各自的诊...肾母细胞瘤(Wilms tumor,WT)是儿童最常见的肾恶性肿瘤。COG(Children’s Oncology Group,儿童肿瘤协作组)和SIOP(International Society of Paediatric Oncology,国际儿童肿瘤学会)两大儿童肿瘤研究的国际协作组织分别形成了各自的诊疗方案:COG推荐手术优先,然后行化疗与放射治疗等辅助治疗;SIOP强调术前化疗,然后行手术与放射治疗。目前该两大协作组发布的肾母细胞瘤患者总体生存率均达到90%。中国抗癌协会小儿肿瘤专业委员会及中华医学会小儿外科学分会泌尿外科学组在COG和SIOP方案的基础上也先后发表了诊疗建议和共识。本文重点介绍COG方案和SIOP方案的异同,主要包括组织学分类、临床分期、危险度分组、治疗方案以及复发后治疗等方面;进而解读中国儿童肾母细胞瘤诊疗共识,分析中国的诊疗现状,供广大临床医师参考和借鉴。展开更多
目的基于高通量RNA测序技术和生物信息学分析鉴定肾母细胞瘤(wilms tumor,WT)中潜在的circRNA-miRNA-mRNA调控网络。方法收集2021年6月至2022年3月期间重庆医科大学附属儿童医院泌尿外科的WT患者的肿瘤切除组织共25例。RNA测序技术分析...目的基于高通量RNA测序技术和生物信息学分析鉴定肾母细胞瘤(wilms tumor,WT)中潜在的circRNA-miRNA-mRNA调控网络。方法收集2021年6月至2022年3月期间重庆医科大学附属儿童医院泌尿外科的WT患者的肿瘤切除组织共25例。RNA测序技术分析4对配对的肿瘤和癌旁正常组织的circRNA(DEcircRNA)和mRNA(DEmRNA)差异表达谱。对差异分子进行基于miRanda的miRNA靶点预测,基于相同位点和协同表达建立完整的circRNA-miRNA-mRNA调控网络。最后验证关键环状RNA的功能。结果筛选出314个DEcircRNAs和1612个DEmRNAs。通过RT-qPCR在组织中随机选择12个circRNA进行表达量验证,其中7个circRNA显示出与测序结果一致的趋势。基因本体论(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析提示DEcircRNA的亲本基因和DEmRNA主要富集到细胞增殖、癌症和多种代谢相关通路中。利用miRanda预测工具识别DEcircRNA和DEmRNA的共同miRNA靶点,以此构建完整的circRNA-miRNA-mRNA网络。基因集富集分析(Gene Set Enrichment Analysis,GSEA)提示ceRNA网络中的靶基因与细胞周期和免疫应答有关。为了鉴定关键的下游靶点,基于STRING数据库建立了127个ceRNA网络中靶基因的PPI(protein-protein interaction)网络,并进一步确定了前10个HUB基因。其中,4个HUB基因(TP53、KANK3、LLGL1和TOPA3)被证实与WT的预后密切相关。基于具有预后意义的关键靶基因构建circRNA作用的调控子网络。最后,通过CCK-8、划痕、Transwell和流式细胞实验,发现沉默子网络中的circRNA(hsa_circ_0009035)抑制了WT细胞的增殖、迁移、侵袭和细胞周期分布(P<0.05)。结论本研究展示了WT中circRNAs的全面表达,构建了潜在的circRNA相关的ceRNA调控网络,深入阐释了WT发生发展的调控机制。展开更多
文摘Nephroblastoma is one of the most common causes of abdominal-pelvic mass in children. It still raises a diagnostic problem because of the wide variety of causes of abdominal-pelvic mass but also because of the delays in consultation in sub-Saharan black Africa and consequently the discovery of the mass at a very late and sometimes metastatic stage. Yet nephroblastoma is a very chemo-sensitive malignancy requiring diagnosis at an early stage;a procedure in which medical imaging is essential. We report the case of a large abdominal-pelvic mass in a three-year-old girl in whom the abdominal-pelvic CT allowed to diagnose nephroblastoma with liver metastases. Our objective is to demonstrate the contribution of computed tomography in the diagnosis of large abdominal-pelvic mass of the child and discuss other causes of abdominal-pelvic mass of the child.
文摘The overall frequency of WT1 gene alterations in Wilms tumor is still unclear in Taiwan. Here we conducted molecular genetic analysis of the WT1 gene in Taiwan Residents patients with Wilms tumor. Polymerase chain reaction and direct sequencing were performed on DNA samples from blood and paraffin-embedded tumor specimens. A constitutional mutation in the WT1 gene was found in one DNA sample from peripheral blood lymphocytes. The remaining DNA samples from peripheral blood lymphocytes and paraffin-embedded tumor specimens were tested negative for both constitutional mutations and somatic mutations. Thus, mutations at other Wilms tumor loci may play an important role in Wilms tumor development.
基金Supported by National Natural Science Foundation of China(No.81403223)Zhejiang Provincial Natural Science Foundation of China(No.LQ14H290003)Science and Technology Foundation of Zhejiang Province(No.2015C33173)
文摘Objective To explore the effects of bufalin on inhibiting proliferation, up-regulating methylation of Wilm’ tumor 1 gene (WT1) as well as its possible mechanisms in human erythroid leukemic (HEL) cells. Methods The HEL cells were treated with bufalin at various concentrations to observe cellular morphology, proliferation assay and cell cycle. The mRNA and protein expression levels of WT1 were detected by reverse transcription polymerase chain reaction (RT-PCR), Western blot and immunocytochemistry, DNA methylation of WT1 and protein expression levels of DNA methyltransferase 3a (DNMT3a) and DNMT3b were analyzed by methylation-specific PCR, and Western blot respectively. Results The bufalin was effective to inhibit proliferation of HEL cells in a dose-dependent manner, their suppression rates were from 23.4%±2.1% to 87.2%±5.4% with an half maximal inhibit concentration (IC<sub>50</sub>) of 0.046 μmol/L. Typical apoptosis morphology was observed in bufalin-treated HEL cells. The proliferation index of cell cycle decreased from 76.4%±1.9% to 49.7%±1.3%. The expression levels of WT1 mRNA and its protein reduced gradually with increasing doses of bufalin, meanwhile, the methylation status of WT1 gene changed from unmethylated into partially or totally methylated. While, the expression levels of DNMT3a and DNMT3b protein gradually increased by bufalin treatment in a dose-dependent manner. Conclusions Bufalin can not only significantly inhibit the proliferation of HEL cells and arrest cell cycle at G<sub>0</sub>/G<sub>1</sub> phase, but also induce cellular apoptosis and down-regulate the expression level of WT1. Our results provide the evidence of bufalin for anti-leukemia, its mechanism may involve in increasing WT1 methylation status which is related to the up-regulation of DNMT3a and DNMT3b proteins in erythroid leukemic HEL cells.
文摘肾母细胞瘤(Wilms tumor,WT)是儿童最常见的肾恶性肿瘤。COG(Children’s Oncology Group,儿童肿瘤协作组)和SIOP(International Society of Paediatric Oncology,国际儿童肿瘤学会)两大儿童肿瘤研究的国际协作组织分别形成了各自的诊疗方案:COG推荐手术优先,然后行化疗与放射治疗等辅助治疗;SIOP强调术前化疗,然后行手术与放射治疗。目前该两大协作组发布的肾母细胞瘤患者总体生存率均达到90%。中国抗癌协会小儿肿瘤专业委员会及中华医学会小儿外科学分会泌尿外科学组在COG和SIOP方案的基础上也先后发表了诊疗建议和共识。本文重点介绍COG方案和SIOP方案的异同,主要包括组织学分类、临床分期、危险度分组、治疗方案以及复发后治疗等方面;进而解读中国儿童肾母细胞瘤诊疗共识,分析中国的诊疗现状,供广大临床医师参考和借鉴。
文摘目的基于高通量RNA测序技术和生物信息学分析鉴定肾母细胞瘤(wilms tumor,WT)中潜在的circRNA-miRNA-mRNA调控网络。方法收集2021年6月至2022年3月期间重庆医科大学附属儿童医院泌尿外科的WT患者的肿瘤切除组织共25例。RNA测序技术分析4对配对的肿瘤和癌旁正常组织的circRNA(DEcircRNA)和mRNA(DEmRNA)差异表达谱。对差异分子进行基于miRanda的miRNA靶点预测,基于相同位点和协同表达建立完整的circRNA-miRNA-mRNA调控网络。最后验证关键环状RNA的功能。结果筛选出314个DEcircRNAs和1612个DEmRNAs。通过RT-qPCR在组织中随机选择12个circRNA进行表达量验证,其中7个circRNA显示出与测序结果一致的趋势。基因本体论(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析提示DEcircRNA的亲本基因和DEmRNA主要富集到细胞增殖、癌症和多种代谢相关通路中。利用miRanda预测工具识别DEcircRNA和DEmRNA的共同miRNA靶点,以此构建完整的circRNA-miRNA-mRNA网络。基因集富集分析(Gene Set Enrichment Analysis,GSEA)提示ceRNA网络中的靶基因与细胞周期和免疫应答有关。为了鉴定关键的下游靶点,基于STRING数据库建立了127个ceRNA网络中靶基因的PPI(protein-protein interaction)网络,并进一步确定了前10个HUB基因。其中,4个HUB基因(TP53、KANK3、LLGL1和TOPA3)被证实与WT的预后密切相关。基于具有预后意义的关键靶基因构建circRNA作用的调控子网络。最后,通过CCK-8、划痕、Transwell和流式细胞实验,发现沉默子网络中的circRNA(hsa_circ_0009035)抑制了WT细胞的增殖、迁移、侵袭和细胞周期分布(P<0.05)。结论本研究展示了WT中circRNAs的全面表达,构建了潜在的circRNA相关的ceRNA调控网络,深入阐释了WT发生发展的调控机制。
