目的:讨论Rspo3基因在乳腺癌组织中的表达情况,启动子甲基化水平及意义。方法:通过The Human Protein Atlas分析Rspo3蛋白在乳腺癌组织中的表达情况;利用Oncomine、GEPIA、UALCAN数据库分析Rspo3 DNA、mRNA在乳腺癌组织中的表达情况;利...目的:讨论Rspo3基因在乳腺癌组织中的表达情况,启动子甲基化水平及意义。方法:通过The Human Protein Atlas分析Rspo3蛋白在乳腺癌组织中的表达情况;利用Oncomine、GEPIA、UALCAN数据库分析Rspo3 DNA、mRNA在乳腺癌组织中的表达情况;利用MethHC和UALCAN数据库分析Rspo3基因启动子区域甲基化水平;利用String-DB分析与Rspo3相互作用的蛋白。结果:在蛋白水平上,Rspo3蛋白在乳腺癌组织中较正常乳腺组织低表达,在DNA、mRNA水平上,Rspo3在乳腺癌组织中较正常乳腺组织显著低表达,且其启动子甲基化水平则显著增高。与Rspo3相关蛋白有LGR(4~6)、UBA52、UBB、UBC、Rspo1、Rspo4、ZNRF3、RNF43,可能调节乳腺癌中Wnt/β-catenin信号传导通路的激活。结论:基于现有的肿瘤数据库进行生物信息学分析,可以发现Rspo3在乳腺癌组织中呈低表达,其启动子甲基化水平则表达增高。这为深入研究乳腺癌,并发现新的乳腺肿瘤标志物提供了大样本数据支持。展开更多
R-spondins are secretory proteins localized in the endoplasmic reticulum and Golgi bodies and are processed through the secretory pathway.Among the R-spondin family,RSPO2 has emanated as a novel regulator of Wnt signa...R-spondins are secretory proteins localized in the endoplasmic reticulum and Golgi bodies and are processed through the secretory pathway.Among the R-spondin family,RSPO2 has emanated as a novel regulator of Wnt signaling,which has now been acknowledged in numerous in vitro and in vivo studies.Cancer is an abnormal growth of cells that proliferates and spreads uncontrollably due to the accumulation of genetic and epigenetic factors that constitutively activate Wnt signaling in various types of cancer.Colorectal cancer (CRC) begins when cells in the colon and rectum follow an indefinite pattern of division due to aberrant Wnt activation as one of the key hallmarks.Decades-long progress in research on R-spondins has demonstrated their oncogenic function in distinct cancer types,particularly CRC.As a critical regulator of the Wnt pathway,it modulates several phenotypes of cells,such as cell proliferation,invasion,migration,and cancer stem cell properties.Recently,RSPO mutations,gene rearrangements,fusions,copy number alterations,and altered gene expression have also been identified in a variety of cancers,including CRC.In this review,we addressed the recent updates regarding the recurrently altered R-spondins with special emphasis on the RSPO2 gene and its involvement in potentiating Wnt signaling in CRC.In addition to the compelling physiological and biological roles in cellular fate and regulation,we propose that RSPO2 would be valuable as a potential biomarker for prognostic,diagnostic,and therapeutic use in CRC.展开更多
为探究特异性顶部盘状底板反应蛋白2(RSPO2)在2种绵羊皮肤中的差异表达及其对毛囊生长相关基因表达的影响,以美利奴羊和小尾寒羊背部皮肤为材料,利用免疫组织化学、实时荧光定量PCR技术和Western-blot技术检测RSPO2在美利奴羊和小尾寒...为探究特异性顶部盘状底板反应蛋白2(RSPO2)在2种绵羊皮肤中的差异表达及其对毛囊生长相关基因表达的影响,以美利奴羊和小尾寒羊背部皮肤为材料,利用免疫组织化学、实时荧光定量PCR技术和Western-blot技术检测RSPO2在美利奴羊和小尾寒羊背部皮肤毛囊中的表达与定位;在皮肤成纤维细胞中过表达和干扰RSPO2,用实时荧光定量PCR和Western-blot检测毛囊生长相关基因及增殖相关基因的表达。结果显示,RSPO2 m RNA和蛋白在美利奴羊和小尾寒羊中均有表达,且美利奴羊中的表达量极显著高于小尾寒羊(P<0.01)。在成纤维细胞中转染过表达RSPO2载体后,β-连环蛋白(β-catenin)和Lef1 m RNA和蛋白的表达量极显著升高(P<0.01),BMP4和TGF-β1 m RNA和蛋白的表达量极显著降低(P<0.01);在成纤维细胞中转染干扰RSPO2载体后,β-catenin和Lef1 m RNA和蛋白的表达量极显著降低(P<0.01),BMP4和TGF-β1 m RNA和蛋白的表达量极显著升高(P<0.01)。同时,试验证实RSPO2可以促进成纤维细胞的增殖。结果表明,RSPO2对绵羊毛囊生长有一定的作用,可能是通过促进成纤维细胞增殖和调控参与毛囊生长相关基因的表达来实现的。展开更多
Objective: The aim of this study was to investigate the effect and possible mechanism of action of roof plate-specific spondin1 (Rspo1) in the apoptosis of rat bone marrow mesenchymal stem cells (BMSCs). Methods: Oste...Objective: The aim of this study was to investigate the effect and possible mechanism of action of roof plate-specific spondin1 (Rspo1) in the apoptosis of rat bone marrow mesenchymal stem cells (BMSCs). Methods: Osteogenic and adipogenic differentiation of BMSCs was identified by Alizarin Red and Oil Red O staining, respectively. BMSC surface markers (cluster of differentiation 29 [CD29], CD90, and CD45) were detected using flow cytometry. BMSCs were transfected with an adenoviral vector encoding Rspo1 (BMSCs-Rspo1 group). The expression levels of Rspo1 gene and Rspo1 protein in the BMSCs-Rspo1 group and the two control groups (untransfected BMSCs group and BMSCs-green fluorescent protein [GFP] group) were analyzed and compared by quantitative polymerase chain reaction and Western blot. The occurrence of apoptosis in the three groups was detected by flow cytometry and acridine orange-ethidium bromide (AO-EB) double dyeing. The activity of the Wnt/β-catenin signaling pathway was evaluated by measuring the expression levels of the key proteins of the pathway (β-catenin, c-Jun N-terminal kinase [JNK], and phospho-JNK). Results: Osteogenic and adipogenic differentiation was confirmed in cultured BMSCs by the positive expression of CD29 and CD90 and the negative expression of CD45. Significantly increased expression levels of Rspo1 protein in the BMSCs-Rspo1 group compared to those in the BMSCs (0.60 ± 0.05 vs. 0.13 ± 0.02;t=95.007, P=0.001) and BMSCs-GFP groups (0.60 ± 0.05 vs. 0.10 ± 0.02;t=104.842, P=0.001) were observed. The apoptotic rate was significantly lower in the BMSCs-Rspo1 group compared with those in the BMSCs group ([24.06 ± 2.37]% vs.[40.87 ± 2.82]%;t =49.872, P =0.002) and the BMSCs-GFP group ([24.06 ± 2.37]% vs.[42.34 ± 0.26]%;t =62.358, P =0.001). In addition, compared to the BMSCs group, the protein expression levels of β-catenin (2.67 ± 0.19 vs. 1.14 ± 0.14;t =-9.217, P =0.000) and JNK (1.87 ± 0.17 vs. 0.61 ± 0.07;t =-22.289, P =0.000) were increased in the BMSCs-Rspo1 group. Compared to the BMSCs-GFP group, the protein expression levels of β-catenin (2.67 ± 0.19 vs. 1.44 ± 0.14;t =-5.692, P =0.000) and JNK (1.87 ± 0.17 vs. 0.53 ± 0.06;t =-10.589, P =0.000) were also upregulated in the BMSCs-Rspo1 group. Moreover, the protein expression levels of phospho-JNK were increased in the BMSCs-Rspo1 group compared to those in the BMSCs group (1.89 ± 0.10 vs. 0.63 ± 0.09;t =-8.975, P =0.001) and the BMSCs-GFP group (1.89 ± 0.10 vs. 0.69 ± 0.08;t =-9.483, P =0.001). Conclusion: The Wnt/β-catenin pathway could play a vital role in the Rspo1-mediated inhibition of apoptosis in BMSCs.展开更多
文摘目的:讨论Rspo3基因在乳腺癌组织中的表达情况,启动子甲基化水平及意义。方法:通过The Human Protein Atlas分析Rspo3蛋白在乳腺癌组织中的表达情况;利用Oncomine、GEPIA、UALCAN数据库分析Rspo3 DNA、mRNA在乳腺癌组织中的表达情况;利用MethHC和UALCAN数据库分析Rspo3基因启动子区域甲基化水平;利用String-DB分析与Rspo3相互作用的蛋白。结果:在蛋白水平上,Rspo3蛋白在乳腺癌组织中较正常乳腺组织低表达,在DNA、mRNA水平上,Rspo3在乳腺癌组织中较正常乳腺组织显著低表达,且其启动子甲基化水平则显著增高。与Rspo3相关蛋白有LGR(4~6)、UBA52、UBB、UBC、Rspo1、Rspo4、ZNRF3、RNF43,可能调节乳腺癌中Wnt/β-catenin信号传导通路的激活。结论:基于现有的肿瘤数据库进行生物信息学分析,可以发现Rspo3在乳腺癌组织中呈低表达,其启动子甲基化水平则表达增高。这为深入研究乳腺癌,并发现新的乳腺肿瘤标志物提供了大样本数据支持。
文摘R-spondins are secretory proteins localized in the endoplasmic reticulum and Golgi bodies and are processed through the secretory pathway.Among the R-spondin family,RSPO2 has emanated as a novel regulator of Wnt signaling,which has now been acknowledged in numerous in vitro and in vivo studies.Cancer is an abnormal growth of cells that proliferates and spreads uncontrollably due to the accumulation of genetic and epigenetic factors that constitutively activate Wnt signaling in various types of cancer.Colorectal cancer (CRC) begins when cells in the colon and rectum follow an indefinite pattern of division due to aberrant Wnt activation as one of the key hallmarks.Decades-long progress in research on R-spondins has demonstrated their oncogenic function in distinct cancer types,particularly CRC.As a critical regulator of the Wnt pathway,it modulates several phenotypes of cells,such as cell proliferation,invasion,migration,and cancer stem cell properties.Recently,RSPO mutations,gene rearrangements,fusions,copy number alterations,and altered gene expression have also been identified in a variety of cancers,including CRC.In this review,we addressed the recent updates regarding the recurrently altered R-spondins with special emphasis on the RSPO2 gene and its involvement in potentiating Wnt signaling in CRC.In addition to the compelling physiological and biological roles in cellular fate and regulation,we propose that RSPO2 would be valuable as a potential biomarker for prognostic,diagnostic,and therapeutic use in CRC.
文摘为探究特异性顶部盘状底板反应蛋白2(RSPO2)在2种绵羊皮肤中的差异表达及其对毛囊生长相关基因表达的影响,以美利奴羊和小尾寒羊背部皮肤为材料,利用免疫组织化学、实时荧光定量PCR技术和Western-blot技术检测RSPO2在美利奴羊和小尾寒羊背部皮肤毛囊中的表达与定位;在皮肤成纤维细胞中过表达和干扰RSPO2,用实时荧光定量PCR和Western-blot检测毛囊生长相关基因及增殖相关基因的表达。结果显示,RSPO2 m RNA和蛋白在美利奴羊和小尾寒羊中均有表达,且美利奴羊中的表达量极显著高于小尾寒羊(P<0.01)。在成纤维细胞中转染过表达RSPO2载体后,β-连环蛋白(β-catenin)和Lef1 m RNA和蛋白的表达量极显著升高(P<0.01),BMP4和TGF-β1 m RNA和蛋白的表达量极显著降低(P<0.01);在成纤维细胞中转染干扰RSPO2载体后,β-catenin和Lef1 m RNA和蛋白的表达量极显著降低(P<0.01),BMP4和TGF-β1 m RNA和蛋白的表达量极显著升高(P<0.01)。同时,试验证实RSPO2可以促进成纤维细胞的增殖。结果表明,RSPO2对绵羊毛囊生长有一定的作用,可能是通过促进成纤维细胞增殖和调控参与毛囊生长相关基因的表达来实现的。
基金Shanxi Scholarship Council of China (grant 2012-048 awarded to Li-Hong Yang, 2013-Key Project 3 to Jun Xie, 2016-051 to Zhi- Zhen Liu)National Natural Science Foundation Projects [81671462] awarded to Jun Xie+1 种基金Shanxi Province Key Laboratory of Birth Defects and Cell Regeneration and Research Project awarded to Jun Xiethe Fund for Shanxi "T331 Project" Key Subjects Construction awarded to Jun Xie.
文摘Objective: The aim of this study was to investigate the effect and possible mechanism of action of roof plate-specific spondin1 (Rspo1) in the apoptosis of rat bone marrow mesenchymal stem cells (BMSCs). Methods: Osteogenic and adipogenic differentiation of BMSCs was identified by Alizarin Red and Oil Red O staining, respectively. BMSC surface markers (cluster of differentiation 29 [CD29], CD90, and CD45) were detected using flow cytometry. BMSCs were transfected with an adenoviral vector encoding Rspo1 (BMSCs-Rspo1 group). The expression levels of Rspo1 gene and Rspo1 protein in the BMSCs-Rspo1 group and the two control groups (untransfected BMSCs group and BMSCs-green fluorescent protein [GFP] group) were analyzed and compared by quantitative polymerase chain reaction and Western blot. The occurrence of apoptosis in the three groups was detected by flow cytometry and acridine orange-ethidium bromide (AO-EB) double dyeing. The activity of the Wnt/β-catenin signaling pathway was evaluated by measuring the expression levels of the key proteins of the pathway (β-catenin, c-Jun N-terminal kinase [JNK], and phospho-JNK). Results: Osteogenic and adipogenic differentiation was confirmed in cultured BMSCs by the positive expression of CD29 and CD90 and the negative expression of CD45. Significantly increased expression levels of Rspo1 protein in the BMSCs-Rspo1 group compared to those in the BMSCs (0.60 ± 0.05 vs. 0.13 ± 0.02;t=95.007, P=0.001) and BMSCs-GFP groups (0.60 ± 0.05 vs. 0.10 ± 0.02;t=104.842, P=0.001) were observed. The apoptotic rate was significantly lower in the BMSCs-Rspo1 group compared with those in the BMSCs group ([24.06 ± 2.37]% vs.[40.87 ± 2.82]%;t =49.872, P =0.002) and the BMSCs-GFP group ([24.06 ± 2.37]% vs.[42.34 ± 0.26]%;t =62.358, P =0.001). In addition, compared to the BMSCs group, the protein expression levels of β-catenin (2.67 ± 0.19 vs. 1.14 ± 0.14;t =-9.217, P =0.000) and JNK (1.87 ± 0.17 vs. 0.61 ± 0.07;t =-22.289, P =0.000) were increased in the BMSCs-Rspo1 group. Compared to the BMSCs-GFP group, the protein expression levels of β-catenin (2.67 ± 0.19 vs. 1.44 ± 0.14;t =-5.692, P =0.000) and JNK (1.87 ± 0.17 vs. 0.53 ± 0.06;t =-10.589, P =0.000) were also upregulated in the BMSCs-Rspo1 group. Moreover, the protein expression levels of phospho-JNK were increased in the BMSCs-Rspo1 group compared to those in the BMSCs group (1.89 ± 0.10 vs. 0.63 ± 0.09;t =-8.975, P =0.001) and the BMSCs-GFP group (1.89 ± 0.10 vs. 0.69 ± 0.08;t =-9.483, P =0.001). Conclusion: The Wnt/β-catenin pathway could play a vital role in the Rspo1-mediated inhibition of apoptosis in BMSCs.
