Spleen tyrosine kinase (SYK),a non-receptor tyrosine kinase,is expressed in most hematopoietic cells and non-hematopoietic cells and play a crucial role in both immune and non-immune biological responses.SYK mediate d...Spleen tyrosine kinase (SYK),a non-receptor tyrosine kinase,is expressed in most hematopoietic cells and non-hematopoietic cells and play a crucial role in both immune and non-immune biological responses.SYK mediate diverse cellular responses via an immune-receptor tyrosine-based activation motifs (ITAMs)-dependent signalling pathways,ITAMs-independent and ITAMs-semidependent signalling pathways.In liver,SYK expression has been observed in parenchymal (hepatocytes) and non-parenchymal cells (hepatic stellate cells and Kupffer cells) and found to be positively correlated with the disease severity.The implication of SYK pathway has been reported in different liver diseases including liver fibrosis,viral hepatitis,alcoholic liver disease,non-alcoholic steatohepatitis and hepatocellular carcinoma.Antagonism of SYK pathway using kinase inhibitors have shown to attenuate the progression of liver diseases thereby suggesting SYK as a highly promising therapeutic target.This review summarizes the current understanding of SYK and its therapeutic implication in liver diseases.展开更多
背景与目的:全长型脾脏酪氨酸激酶[full-length form of spleen tyrosine kinase,Syk(L)]在乳腺癌中具有抑癌功能,可移位到细胞核内,作为转录抑制因子调节基因的转录。本研究拟探讨Syk(L)行使转录抑制功能的具体分子生物学机制。方法:将...背景与目的:全长型脾脏酪氨酸激酶[full-length form of spleen tyrosine kinase,Syk(L)]在乳腺癌中具有抑癌功能,可移位到细胞核内,作为转录抑制因子调节基因的转录。本研究拟探讨Syk(L)行使转录抑制功能的具体分子生物学机制。方法:将pFLAG-CMV-Syk(L)转染入人胚肾细胞HEK293中,采用免疫沉淀方法检测组蛋白去乙酰化酶(histone deacetylases,HDACs)与外源性Syk(L)的结合。采用免疫沉淀方法检测乳腺癌细胞MB468中内源性Syk(L)与HDAC1的结合。构建带有FLAG标志的Syk(L)各个功能域质粒,并将其转染入HEK293细胞中,采用免疫沉淀方法检测HDAC1与Syk(L)各个功能域之间的结合。采用HDAC活力检测系统检测Syk(L)免疫沉淀复合物中的HDAC活性。结果:HEK293细胞中外源性Syk(L)可与HDAC1、3、6、7结合,Syk(L)通过SH2功能域和KD功能域与HDAC1相互结合。乳腺癌细胞MB468中内源性Syk(L)与HDAC1可相互结合。Syk(L)免疫沉淀复合物有明显的HDAC活性。结论:在乳腺癌中,抑癌基因Syk(L)可能通过与HDAC形成复合物调节基因的转录。展开更多
目的探讨天花粉蛋白通过全长型脾络氨酸激酶[full-length form of spleen tyrosine kinase,Syk(L)]基因去甲基化途径恢复Syk(L)表达对喉癌Hep-2细胞增殖、侵袭迁移的的抑制作用。方法采用CCK-8法及Transwell实验检测不同浓度天花粉在不...目的探讨天花粉蛋白通过全长型脾络氨酸激酶[full-length form of spleen tyrosine kinase,Syk(L)]基因去甲基化途径恢复Syk(L)表达对喉癌Hep-2细胞增殖、侵袭迁移的的抑制作用。方法采用CCK-8法及Transwell实验检测不同浓度天花粉在不同时间点对Hep-2细胞增殖及处理72h后细胞侵袭迁移能力的影响;采用MS-HRM法、Q-RT-PCR及Western blot法检测不同浓度天花粉蛋白对Hep-2细胞Syk(L)基因甲基化水平及其表达的影响。结果 CCK-8结果显示,天花粉蛋白对喉癌Hep-2细胞的增殖有抑制作用,且呈浓度、结果时间依赖性;Transwell结果显示,天花粉蛋白能显著抑制Hep-2细胞的侵袭迁移能力;MS-HRM结果显示,天花粉蛋白可以逆转Hep-2细胞Sky(L)基因的高甲基化状态,恢复Syk(L)m RNA及蛋白的表达。结论天花粉蛋白通过逆转Syk(L)基因启动子Cp G岛的甲基化状态,使喉癌Hep-2细胞中Syk(L)基因表达活化,恢复Syk m RNA及蛋白的表达,最终发挥抑癌基因的作用抑制Hep-2细胞恶性生物学行为,具有开发并成为抗肿瘤药物的潜在价值。展开更多
基金Supported by the Endowment Fund for the Education Republic of Indonesia(Lembaga Pengelola Dana Pendidikan/LPDP RI)No.44/LPDP/2015
文摘Spleen tyrosine kinase (SYK),a non-receptor tyrosine kinase,is expressed in most hematopoietic cells and non-hematopoietic cells and play a crucial role in both immune and non-immune biological responses.SYK mediate diverse cellular responses via an immune-receptor tyrosine-based activation motifs (ITAMs)-dependent signalling pathways,ITAMs-independent and ITAMs-semidependent signalling pathways.In liver,SYK expression has been observed in parenchymal (hepatocytes) and non-parenchymal cells (hepatic stellate cells and Kupffer cells) and found to be positively correlated with the disease severity.The implication of SYK pathway has been reported in different liver diseases including liver fibrosis,viral hepatitis,alcoholic liver disease,non-alcoholic steatohepatitis and hepatocellular carcinoma.Antagonism of SYK pathway using kinase inhibitors have shown to attenuate the progression of liver diseases thereby suggesting SYK as a highly promising therapeutic target.This review summarizes the current understanding of SYK and its therapeutic implication in liver diseases.
文摘背景与目的:全长型脾脏酪氨酸激酶[full-length form of spleen tyrosine kinase,Syk(L)]在乳腺癌中具有抑癌功能,可移位到细胞核内,作为转录抑制因子调节基因的转录。本研究拟探讨Syk(L)行使转录抑制功能的具体分子生物学机制。方法:将pFLAG-CMV-Syk(L)转染入人胚肾细胞HEK293中,采用免疫沉淀方法检测组蛋白去乙酰化酶(histone deacetylases,HDACs)与外源性Syk(L)的结合。采用免疫沉淀方法检测乳腺癌细胞MB468中内源性Syk(L)与HDAC1的结合。构建带有FLAG标志的Syk(L)各个功能域质粒,并将其转染入HEK293细胞中,采用免疫沉淀方法检测HDAC1与Syk(L)各个功能域之间的结合。采用HDAC活力检测系统检测Syk(L)免疫沉淀复合物中的HDAC活性。结果:HEK293细胞中外源性Syk(L)可与HDAC1、3、6、7结合,Syk(L)通过SH2功能域和KD功能域与HDAC1相互结合。乳腺癌细胞MB468中内源性Syk(L)与HDAC1可相互结合。Syk(L)免疫沉淀复合物有明显的HDAC活性。结论:在乳腺癌中,抑癌基因Syk(L)可能通过与HDAC形成复合物调节基因的转录。
文摘目的探讨天花粉蛋白通过全长型脾络氨酸激酶[full-length form of spleen tyrosine kinase,Syk(L)]基因去甲基化途径恢复Syk(L)表达对喉癌Hep-2细胞增殖、侵袭迁移的的抑制作用。方法采用CCK-8法及Transwell实验检测不同浓度天花粉在不同时间点对Hep-2细胞增殖及处理72h后细胞侵袭迁移能力的影响;采用MS-HRM法、Q-RT-PCR及Western blot法检测不同浓度天花粉蛋白对Hep-2细胞Syk(L)基因甲基化水平及其表达的影响。结果 CCK-8结果显示,天花粉蛋白对喉癌Hep-2细胞的增殖有抑制作用,且呈浓度、结果时间依赖性;Transwell结果显示,天花粉蛋白能显著抑制Hep-2细胞的侵袭迁移能力;MS-HRM结果显示,天花粉蛋白可以逆转Hep-2细胞Sky(L)基因的高甲基化状态,恢复Syk(L)m RNA及蛋白的表达。结论天花粉蛋白通过逆转Syk(L)基因启动子Cp G岛的甲基化状态,使喉癌Hep-2细胞中Syk(L)基因表达活化,恢复Syk m RNA及蛋白的表达,最终发挥抑癌基因的作用抑制Hep-2细胞恶性生物学行为,具有开发并成为抗肿瘤药物的潜在价值。
