目的构建含有全长WTX(Wilms tumor gene on the X chromosome)目的基因的表达载体,建立稳定表达WTX基因的胃癌BGC-823/WTX-EGFP细胞系,为WTX基因功能研究提供保障。方法以人胚肾293FT细胞为模板,经PCR扩增获得全长WTX cDNA序列,将全长WT...目的构建含有全长WTX(Wilms tumor gene on the X chromosome)目的基因的表达载体,建立稳定表达WTX基因的胃癌BGC-823/WTX-EGFP细胞系,为WTX基因功能研究提供保障。方法以人胚肾293FT细胞为模板,经PCR扩增获得全长WTX cDNA序列,将全长WTX目的基因序列克隆入带有绿色荧光蛋白报告基因的pEGFP-N1表达载体,建立pEGFP-WTX表达载体。经限制性内切酶酶切鉴定、测序并转染293FT细胞观察报告基因表达情况,判断pEGFP-WTX表达载体构建是否成功。构建成功的pEGFP-WTX表达载体转染人胃癌BGC-823细胞,建立稳定表达WTX的BGC-823/WTX-EGFP细胞。qRT-PCR及免疫组化检测转染细胞WTX表达。结果 PCR方法从人胚肾293FT细胞扩增获得全长WTX cDNA,经测序鉴定序列完全正确;pEGFP-WTX质粒DNA经酶切鉴定片段大小正确、转染294FT细胞可见清晰的绿色荧光表达。BGC-823/WTX-EGFP细胞能够稳定表达WTX。结论本研究成功构建了含有全长WTX cDNA的表达载体系统,建立了稳定表达WTX的BGC-823/WTX-EGFP细胞,为WTX基因功能研究奠定了良好基础。展开更多
Wilms tumor(WT) is one of the most common renal malignancies in children.Although several genetic loci such as the WT1,WT2,p53 and β-catenin genes have been considered to be associated with WT,the causes of the tumor...Wilms tumor(WT) is one of the most common renal malignancies in children.Although several genetic loci such as the WT1,WT2,p53 and β-catenin genes have been considered to be associated with WT,the causes of the tumor are still unknown.Recently the US researchers have identified a new tumor suppressor gene that is mutated in WT.The biological function of the protein encoded by WTX is yet unknown,however,the gene’s location in the X chromosome is of particular interest.This review highlights the current study of the gene mutated in Wilms tumor.展开更多
肿瘤的发生发展与癌基因的活化及抑癌基因的失活密切相关早已得到公认.近年来,多种抑癌基因陆续被发现并明确其作用机制.2007年Rivera等在研究Wilms’瘤时发现了一个新的抑癌基因WTX(Wilms tumor gene on the X chromosome),该基因是第...肿瘤的发生发展与癌基因的活化及抑癌基因的失活密切相关早已得到公认.近年来,多种抑癌基因陆续被发现并明确其作用机制.2007年Rivera等在研究Wilms’瘤时发现了一个新的抑癌基因WTX(Wilms tumor gene on the X chromosome),该基因是第一次在X染色体上发现的直接作为肿瘤抑制因子的基因,并首次在《Science》上报道.展开更多
Osteopathia striata with cranial sclerosis(OSCS, OMIM#300373) is an X-linked dominant sclerosing bone dysplasia that shows a distinct phenotype in females and males. In 2009, Zandra Jenkins et al found that germline m...Osteopathia striata with cranial sclerosis(OSCS, OMIM#300373) is an X-linked dominant sclerosing bone dysplasia that shows a distinct phenotype in females and males. In 2009, Zandra Jenkins et al found that germline mutations in the FAM123 B /WTX /AMER1 gene, mapped to chromosome Xq11.2, cause both the familial and sporadic forms of OSCS. Intriguingly, the WTX gene was already known as a putative tumor suppressor gene, since in 2007 Rivera et al had reported inactivating WTX mutations in Wilms' tumor(WT), the most frequent renal tumor of childhood. Here we review the heterogeneous clinical presentation of OSCS patients and the involvement of WTX anomalies in OSCS and in WT.展开更多
Background Wilms＇ tumor （nephroblastoma） is the most common pediatric kidney cancer. Only one Wilms＇ tumor gene is known, WT1 at 11p13, which is mutated in 5%-10% of Wilms＇ tumors. Recently, mutations were report...Background Wilms＇ tumor （nephroblastoma） is the most common pediatric kidney cancer. Only one Wilms＇ tumor gene is known, WT1 at 11p13, which is mutated in 5%-10% of Wilms＇ tumors. Recently, mutations were reported in WTX at Xq11.1 in Wilms＇ tumors. This study investigated the mutation proportion, type, and distribution in WTX and WT1 in children with Wilms＇ tumor. The role of WTX/WT1 in the development of Wilms＇ tumor, and the relationship between clinical phenotype and genotype, were also studied. Methods Wilms＇ tumor specimens （blood samples from 70 patients and tumor tissue samples from 52 patients） were used. A long fragment of WTXand 10 exons and intron sequences of WT1 were amplified by polymerase chain reaction （PCR） from extracted genomic DNA and sequenced. A chi-square test compared the difference between the W-/-X mutation group and the no mutation group. The relationship between the mutations and clinical phenotype was analyzed. Results W7X mutations were found in 5/52 tumor tissues and in 2/70 peripheral blood samples （five cases in total, all point mutations）. Two patients had a WTX mutation in both samples. WT1 mutations were found in 2/52 tumor tissues and in 4/70 peripheral blood samples （five cases in total, all point mutations）. One patient had a WT1 mutation in both samples. Ten cases had WTX or WT1 mutation （19.2% of Wilms＇ tumors）. No overlapping WTX and WTI mutations were found. No significant differences in clinical parameters were found between patients with and without a W7X mutation. Conclusions WTX mutations occur early in Wilms＇ tumor development, but at a low proportion. There was no evidence that WTX is the main cause of Wilms＇ tumor. Clinical parameters of patients with WTX mutations are not related to the mutation, indicating a limited impact of WTX on tumor progression. WTX and WT1 mutations occur independently, suggesting a relationship between their gene products.展开更多
基金Supported by National Natural Science Foundation of China(81071989,81272760)Natural Science Foundation of Guangdong Province(S2011010004178)Science and Technology Program of Guangdong Province(c1221020700008)~~
文摘目的构建含有全长WTX(Wilms tumor gene on the X chromosome)目的基因的表达载体,建立稳定表达WTX基因的胃癌BGC-823/WTX-EGFP细胞系,为WTX基因功能研究提供保障。方法以人胚肾293FT细胞为模板,经PCR扩增获得全长WTX cDNA序列,将全长WTX目的基因序列克隆入带有绿色荧光蛋白报告基因的pEGFP-N1表达载体,建立pEGFP-WTX表达载体。经限制性内切酶酶切鉴定、测序并转染293FT细胞观察报告基因表达情况,判断pEGFP-WTX表达载体构建是否成功。构建成功的pEGFP-WTX表达载体转染人胃癌BGC-823细胞,建立稳定表达WTX的BGC-823/WTX-EGFP细胞。qRT-PCR及免疫组化检测转染细胞WTX表达。结果 PCR方法从人胚肾293FT细胞扩增获得全长WTX cDNA,经测序鉴定序列完全正确;pEGFP-WTX质粒DNA经酶切鉴定片段大小正确、转染294FT细胞可见清晰的绿色荧光表达。BGC-823/WTX-EGFP细胞能够稳定表达WTX。结论本研究成功构建了含有全长WTX cDNA的表达载体系统,建立了稳定表达WTX的BGC-823/WTX-EGFP细胞,为WTX基因功能研究奠定了良好基础。
文摘Wilms tumor(WT) is one of the most common renal malignancies in children.Although several genetic loci such as the WT1,WT2,p53 and β-catenin genes have been considered to be associated with WT,the causes of the tumor are still unknown.Recently the US researchers have identified a new tumor suppressor gene that is mutated in WT.The biological function of the protein encoded by WTX is yet unknown,however,the gene’s location in the X chromosome is of particular interest.This review highlights the current study of the gene mutated in Wilms tumor.
文摘肿瘤的发生发展与癌基因的活化及抑癌基因的失活密切相关早已得到公认.近年来,多种抑癌基因陆续被发现并明确其作用机制.2007年Rivera等在研究Wilms’瘤时发现了一个新的抑癌基因WTX(Wilms tumor gene on the X chromosome),该基因是第一次在X染色体上发现的直接作为肿瘤抑制因子的基因,并首次在《Science》上报道.
基金Supported by Associazione Bianca Garavaglia,21052 BustoArsizio,Varese,ItalyItalian Association for Cancer Research(AIRC)Fondazione Pierfranco e Luisa Mariani,20129 Milano,Italy
文摘Osteopathia striata with cranial sclerosis(OSCS, OMIM#300373) is an X-linked dominant sclerosing bone dysplasia that shows a distinct phenotype in females and males. In 2009, Zandra Jenkins et al found that germline mutations in the FAM123 B /WTX /AMER1 gene, mapped to chromosome Xq11.2, cause both the familial and sporadic forms of OSCS. Intriguingly, the WTX gene was already known as a putative tumor suppressor gene, since in 2007 Rivera et al had reported inactivating WTX mutations in Wilms' tumor(WT), the most frequent renal tumor of childhood. Here we review the heterogeneous clinical presentation of OSCS patients and the involvement of WTX anomalies in OSCS and in WT.
文摘Background Wilms＇ tumor （nephroblastoma） is the most common pediatric kidney cancer. Only one Wilms＇ tumor gene is known, WT1 at 11p13, which is mutated in 5%-10% of Wilms＇ tumors. Recently, mutations were reported in WTX at Xq11.1 in Wilms＇ tumors. This study investigated the mutation proportion, type, and distribution in WTX and WT1 in children with Wilms＇ tumor. The role of WTX/WT1 in the development of Wilms＇ tumor, and the relationship between clinical phenotype and genotype, were also studied. Methods Wilms＇ tumor specimens （blood samples from 70 patients and tumor tissue samples from 52 patients） were used. A long fragment of WTXand 10 exons and intron sequences of WT1 were amplified by polymerase chain reaction （PCR） from extracted genomic DNA and sequenced. A chi-square test compared the difference between the W-/-X mutation group and the no mutation group. The relationship between the mutations and clinical phenotype was analyzed. Results W7X mutations were found in 5/52 tumor tissues and in 2/70 peripheral blood samples （five cases in total, all point mutations）. Two patients had a WTX mutation in both samples. WT1 mutations were found in 2/52 tumor tissues and in 4/70 peripheral blood samples （five cases in total, all point mutations）. One patient had a WT1 mutation in both samples. Ten cases had WTX or WT1 mutation （19.2% of Wilms＇ tumors）. No overlapping WTX and WTI mutations were found. No significant differences in clinical parameters were found between patients with and without a W7X mutation. Conclusions WTX mutations occur early in Wilms＇ tumor development, but at a low proportion. There was no evidence that WTX is the main cause of Wilms＇ tumor. Clinical parameters of patients with WTX mutations are not related to the mutation, indicating a limited impact of WTX on tumor progression. WTX and WT1 mutations occur independently, suggesting a relationship between their gene products.
