BACKGROUND We present a case of an EWSR1/FUS::NFATC2 rearranged sarcoma in the left forearm and analyze its clinicopathological and molecular features.CASE SUMMARY The patient is a 23-year-old woman.Microscopically,th...BACKGROUND We present a case of an EWSR1/FUS::NFATC2 rearranged sarcoma in the left forearm and analyze its clinicopathological and molecular features.CASE SUMMARY The patient is a 23-year-old woman.Microscopically,the tumor cells were medium-sized round cells arranged in small nests.The cytoplasm was clear,nuclei were relatively uniform,chromatin was dense,nucleoli were visible,and mitotic figures were rare.Immunohistochemically,the tumor cells were positive for Vimentin,INI-1,CD99,NKX2.2,CyclinD1,friend leukaemia virus integration 1,and NKX3.1.Next-generation sequencing revealed the presence of the EWSR1-NFATC2 fusion gene.EWSR1/FUS::NFATC2 rearranged sarcomas are rare and can easily be misdiagnosed.CONCLUSION Clinical imaging,immunohistochemistry,and molecular pathology should be considered to confirm the diagnosis.展开更多
目的构建人T细胞活化核因子C2(NFATc2)基因启动子荧光素酶报告基因载体pGL3-NFATc2-promoter,检测其转录活性,探究二甲双胍和脂多糖对其转录活性的影响。方法利用UCSC网站查找人NFATc2基因的启动子序列并设计上下游引物PCR扩增人NFATc2...目的构建人T细胞活化核因子C2(NFATc2)基因启动子荧光素酶报告基因载体pGL3-NFATc2-promoter,检测其转录活性,探究二甲双胍和脂多糖对其转录活性的影响。方法利用UCSC网站查找人NFATc2基因的启动子序列并设计上下游引物PCR扩增人NFATc2基因启动子片段;用限制性内切酶KpnⅠ和HindⅢ双酶切质粒pGL3-basic,将人NFATc2基因启动子片段插入到pGL3-basic质粒,重组质粒命名为pGL3-NFATc2-promoter。将pGL3-NFATc2-promoter与内参质粒pRL-TK共转染293F细胞,检测其荧光素酶活性。同时构建人NFATc2基因启动子不同片段长度的报告基因载体并进行荧光素酶活性检测,分别给予不同浓度的二甲双胍和脂多糖处理24 h后检测二甲双胍和脂多糖对NFATc2转录活性的影响。进一步突变NFATC2基因启动子上转录因子RUNX2的结合位点探究二甲双胍和脂多糖对NFATc2的转录调控作用。结果研究成功构建了不同片段长度(2170、2077、1802、1651、1083、323 bp)的人NFATC2基因启动子荧光素酶报告基因载体pGL3-NFATc2-promoter,经酶切及测序鉴定完全正确。将不同片段长度的pGL3-NFATc2-promoter转染到293F细胞,发现pGL3-1651 bp具有最高转录活性,荧光素酶活性约为pGL3-2170 bp的3.3倍(1.8433±0.1457 vs 0.5467±0.0850)。中浓度(5 mmol/L)和高浓度(10 mmol/L)的二甲双胍分别上调pGL3-1651 bp的转录活性多达2.5、3倍(1.3467±0.1601 vs 0.8867±0.0321,1.8124±0.2771 vs 0.8867±0.0321)。不同剂量的脂多糖均能上调pGL3-1651 bp的转录活性不低于1.6倍(1.4813±0.0616 vs 0.8867±0.0321)。突变pGL3-1651 bp上的RUNX2结合位点后,二甲双胍和脂多糖上调的pGL3-1651 bp转录活性均受到抑制,转录活性下降(2.1667±0.1527 vs 1.233±0.1155;2.3667±0.2887 vs 1.1333±0.3786)。结论pGL3-NFATc2-promoter在293F细胞中能被转录激活,并证实脂多糖和二甲双胍转录激活pGL3-NFATc2-promoter依赖于转录因RUNX2。展开更多
目的分析口腔鳞状细胞癌(oral squamous cell carcinomas,OSCC)组织中与受体型蛋白酪氨酸磷酸酶K(receptor protein tyrosine phosphatase-K,PTPRK)相关的胞质活化T细胞核因子c2蛋白(nuclear factor of activated T cells c2,NFATc2)的...目的分析口腔鳞状细胞癌(oral squamous cell carcinomas,OSCC)组织中与受体型蛋白酪氨酸磷酸酶K(receptor protein tyrosine phosphatase-K,PTPRK)相关的胞质活化T细胞核因子c2蛋白(nuclear factor of activated T cells c2,NFATc2)的表达和定位,探讨PTPRK抑制OSCC的潜在机制。方法生物信息学预测与PTPRK相互作用的蛋白,免疫组化检测OSCC组织及相应癌旁口腔黏膜组织(对照组)中NFATc2表达及其与PTPRK的关系,免疫荧光染色检测NFATc2在人正常口腔上皮细胞系HIOEC和OSCC细胞系SCC25中的表达和细胞核定位情况,并分析其蛋白表达与OSCC临床病理特征的关系。结果生物信息学预测结果显示在分子通路方面PTPRK可能与NFATc2关联;与对照组比较,NFATc2在OSCC组织中的表达显著升高(P<0.05),且与PTPRK表达呈负相关(P<0.01);细胞免疫荧光染色结果显示,与人正常口腔上皮细胞系HIOEC比较,OSCC细胞系SCC25中NFATc2表达水平升高且表达入核;OSCC中NFATc2表达与临床分期和病理分级相关(P均<0.05)。结论NFATc2表达增加且入核与PTPRK表达减少相关,并参与OSCC的发生、进展。展开更多
基金Supported by The Shenzhen Science and Technology Program,No.JCYJ20220530144407017.
文摘BACKGROUND We present a case of an EWSR1/FUS::NFATC2 rearranged sarcoma in the left forearm and analyze its clinicopathological and molecular features.CASE SUMMARY The patient is a 23-year-old woman.Microscopically,the tumor cells were medium-sized round cells arranged in small nests.The cytoplasm was clear,nuclei were relatively uniform,chromatin was dense,nucleoli were visible,and mitotic figures were rare.Immunohistochemically,the tumor cells were positive for Vimentin,INI-1,CD99,NKX2.2,CyclinD1,friend leukaemia virus integration 1,and NKX3.1.Next-generation sequencing revealed the presence of the EWSR1-NFATC2 fusion gene.EWSR1/FUS::NFATC2 rearranged sarcomas are rare and can easily be misdiagnosed.CONCLUSION Clinical imaging,immunohistochemistry,and molecular pathology should be considered to confirm the diagnosis.
文摘目的构建人T细胞活化核因子C2(NFATc2)基因启动子荧光素酶报告基因载体pGL3-NFATc2-promoter,检测其转录活性,探究二甲双胍和脂多糖对其转录活性的影响。方法利用UCSC网站查找人NFATc2基因的启动子序列并设计上下游引物PCR扩增人NFATc2基因启动子片段;用限制性内切酶KpnⅠ和HindⅢ双酶切质粒pGL3-basic,将人NFATc2基因启动子片段插入到pGL3-basic质粒,重组质粒命名为pGL3-NFATc2-promoter。将pGL3-NFATc2-promoter与内参质粒pRL-TK共转染293F细胞,检测其荧光素酶活性。同时构建人NFATc2基因启动子不同片段长度的报告基因载体并进行荧光素酶活性检测,分别给予不同浓度的二甲双胍和脂多糖处理24 h后检测二甲双胍和脂多糖对NFATc2转录活性的影响。进一步突变NFATC2基因启动子上转录因子RUNX2的结合位点探究二甲双胍和脂多糖对NFATc2的转录调控作用。结果研究成功构建了不同片段长度(2170、2077、1802、1651、1083、323 bp)的人NFATC2基因启动子荧光素酶报告基因载体pGL3-NFATc2-promoter,经酶切及测序鉴定完全正确。将不同片段长度的pGL3-NFATc2-promoter转染到293F细胞,发现pGL3-1651 bp具有最高转录活性,荧光素酶活性约为pGL3-2170 bp的3.3倍(1.8433±0.1457 vs 0.5467±0.0850)。中浓度(5 mmol/L)和高浓度(10 mmol/L)的二甲双胍分别上调pGL3-1651 bp的转录活性多达2.5、3倍(1.3467±0.1601 vs 0.8867±0.0321,1.8124±0.2771 vs 0.8867±0.0321)。不同剂量的脂多糖均能上调pGL3-1651 bp的转录活性不低于1.6倍(1.4813±0.0616 vs 0.8867±0.0321)。突变pGL3-1651 bp上的RUNX2结合位点后,二甲双胍和脂多糖上调的pGL3-1651 bp转录活性均受到抑制,转录活性下降(2.1667±0.1527 vs 1.233±0.1155;2.3667±0.2887 vs 1.1333±0.3786)。结论pGL3-NFATc2-promoter在293F细胞中能被转录激活,并证实脂多糖和二甲双胍转录激活pGL3-NFATc2-promoter依赖于转录因RUNX2。
文摘目的分析口腔鳞状细胞癌(oral squamous cell carcinomas,OSCC)组织中与受体型蛋白酪氨酸磷酸酶K(receptor protein tyrosine phosphatase-K,PTPRK)相关的胞质活化T细胞核因子c2蛋白(nuclear factor of activated T cells c2,NFATc2)的表达和定位,探讨PTPRK抑制OSCC的潜在机制。方法生物信息学预测与PTPRK相互作用的蛋白,免疫组化检测OSCC组织及相应癌旁口腔黏膜组织(对照组)中NFATc2表达及其与PTPRK的关系,免疫荧光染色检测NFATc2在人正常口腔上皮细胞系HIOEC和OSCC细胞系SCC25中的表达和细胞核定位情况,并分析其蛋白表达与OSCC临床病理特征的关系。结果生物信息学预测结果显示在分子通路方面PTPRK可能与NFATc2关联;与对照组比较,NFATc2在OSCC组织中的表达显著升高(P<0.05),且与PTPRK表达呈负相关(P<0.01);细胞免疫荧光染色结果显示,与人正常口腔上皮细胞系HIOEC比较,OSCC细胞系SCC25中NFATc2表达水平升高且表达入核;OSCC中NFATc2表达与临床分期和病理分级相关(P均<0.05)。结论NFATc2表达增加且入核与PTPRK表达减少相关,并参与OSCC的发生、进展。
