脾酪氨酸激酶Syk在食管癌中的表达及对其生物学特征影响的研究
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摘要
目的:恶性肿瘤是严重影响人类生命健康的一类疾病,其发病率呈逐年上升趋势,死亡率居各种疾病之首。肿瘤的发生是一个多因素、多步骤、多基因参与的复杂病理过程,涉及到多种癌基因的激活、抑癌基因的失活和其他调节因子的失调。超过50%的原癌基因和癌基因产物都具有蛋白酪氨酸激酶(protein tyrosine kinases,PTKs)活性。脾酪氨酸激酶(Spleen Tyrosine Kinase,Syk)是一种非受体型蛋白酪氨酸激酶,广泛表达于造血细胞、淋巴细胞、成纤维细胞、血管内皮细胞等细胞,在细胞信号转导中发挥着重要作用。近年的研究认为Syk对多种腺上皮来源的肿瘤生长和转移有抑制作用,但对其在鳞癌中的作用研究较少。本研究拟对Syk在食管鳞状细胞癌组织中的表达情况进行研究,并初步探讨Syk对食管癌细胞生物学性状的影响。
方法:
1采用逆转录-聚合酶链反应(RT-PCR)检测30例食管鳞癌组织、27例食管鳞癌癌旁组织及30例正常食管黏膜组织中Syk mRNA的表达。免疫组化技术法检测39例食管鳞癌组织、27例食管鳞癌癌旁组织及38例正常食管黏膜组织中Syk蛋白的表达。分析Syk蛋白表达与临床病理学指标间的关系。
2用RT-PCR和Western-blot方法检测四种食管癌细胞株(Yes-2、Eca-109、TE-13和TE1)Syk的表达情况。
3给予不同浓度(0、10、50μmol/L)的Syk选择性抑制剂Piceatannol处理Syk阳性表达的TE-1细胞,观察Syk基因的表达变化;通过细胞划痕实验观察处理后细胞迁移能力的改变;采用RT-PCR法分析MMP-2、MMP-9的表达变化。
4 Syk基因甲基化状态检测:提取TE-1和Yes-2细胞DNA,经亚硫酸盐修饰纯化后,行甲基化特异性PCR(methylation-specific PCR, MSP)检测Syk启动子区的甲基化情况。
结果:
1 RT-PCR检测结果显示,在30例食管鳞癌、27例食管鳞癌癌旁组织和30例正常食管黏膜组织中,食管癌组织Syk mRNA表达量高于癌旁和正常黏膜组织,统计学分析表明,三组间Syk mRNA的表达差异无统计学意义。经抗Syk单抗进行免疫组化染色,Syk蛋白阳性反应为棕黄色颗粒,定位于细胞浆。结果显示在食管鳞癌组织中34例Syk蛋白表达阳性(34/39, 87.2%),癌旁组织18例表达阳性(18/27, 66.7%),正常食管黏膜组织中26例表达阳性(26/38, 68.4%),食管癌组织Syk蛋白表达高于食管癌旁组织和正常食管黏膜组织,三组间Syk蛋白表达差异具有统计学意义(P<0.05)。Syk蛋白表达与食管鳞癌患者年龄、性别、肿瘤大小、临床分期差异均无统计学意义(P均>0.05)。
2用RT-PCR和Western-blot方法检测四种不同食管癌细胞株,结果显示Syk基因和蛋白在食管癌细胞株TE1细胞中表达阳性,在Yes-2细胞中表达阴性,Eca-109、TE-13细胞表达为弱阳性。
3 Syk表达阳性的TE-1细胞在给予不同浓度(0、10、50μmol/L)Syk选择性抑制剂Piceatannol处理后,表达逐渐减弱;并且细胞迁移和侵袭能力逐渐恢复;MMP-2和MMP-9的表达逐渐上调。
4 MSP结果显示,TE-1细胞中未发现Syk基因启动子甲基化,Yes-2细胞中发现有Syk基因Syk基因启动子甲基化。
结论:
1 Syk基因和蛋白在食管鳞癌组织中表达增高,但Syk蛋白表达与临床病理学指标无关。
2 Syk表达下调可能参与了食管癌细胞的迁移、侵袭;且其启动子区cpG岛的甲基化参与了食管癌细胞的基因调控。
Objective: Cancer is a class of threatening diseases which has serious impact on health. Morbidity and mortality rate of malignant tumor is increasing in recent years. Tumorigenesis is a multifactor, multistep, polygene involved process that consists of complex pathological changes, such as activation of a variety of oncogenes, inactivation of tumor suppressor genes, and the disorder of other related regulatory factors. More than 50% of proto-oncogene and oncogene proteins have been characterized with the protein- tyrosine kinase (PTKs) activity. Spleen Tyrosine Kinase (Syk) is a non-receptor type of protein- tyrosine kinase that is widely expressed in hematopoietic cells, lymphocyte, fibroblast and vascular endothelial cells. Syk plays an essential role in lymphocyte development and activation. Recent studies have shown that Syk plays an important role in suppressing the growth and metastasis of human glandular epithelium malignant tumor cells with different origins. However, there was few studies paid attention to the expression of Syk in squamous cell carcinoma. In this study, we investigated the gene and protein expression of Syk in human esophageal squamous cell carcinoma (ESCC) and analyzed the effects of Syk on the biocharacteristics of ESCC.
Methods:
1 The expression of Syk gene in human esophageal carcinoma tissues (n=30), peri-cancer tissues (n=27) and normal tunica mucosa esophageal tissues (n=30) were detected by Reverse transcription-polymerase chain reaction (RT-PCR) technique. Immunohistochemistry was used to detect the expression of Syk protein in human esophageal carcinoma tissues (n=30), peri-cancer tissues (n=27) and normal tunica mucosa esophageal tissues (n=30). The relationships between the expression of Syk protein and clinical pathology features were analyzed.
2 RT-PCR and Western-blot was used to measure the expression of Syk gene and Syk protein in esophageal carcinoma cell lines of Yes-2, Eca-109, TE-13 and TE1 cells.
3 Expression of Syk gene was analyzed by RT-PCR, cell migrations were observed by wound healing assay, and expression of MMP-2, MMP-9 was detected by RT-PCR in TE-1 cells after treatment with Piceatannol of different concentrations (0、10、50μmol/L) .
4 The methylation status of the 5’CpG island located in the promoter region of Syk gene in esophageal carcinoma cell lines Yes-2 and TE-1 was detected by methylation specific PCR.
Results:
1 According to the results of RT-PCR, the mRNA expression of the Syk was higher in human esophageal carcinoma lesion tissues than the peri-cancer tissues and the normal esophageal tissues, but among the human esophageal carcinoma lesion tissues, peri-cancer tissues and normal tunica mucosa esophageal tissues the mRNA expression of the Syk had no statistical significance. By immunohistochemical staining with antibody of Sky, it was considered as positive if Syk was expressed in cytoplasm. 34 were Syk protein positive of the human esophageal carcinoma tissues samples (34/39, 87.2%), 18 Syk protein were detected in peri-cancer tissues (18/27, 66.7%) and 26 Syk protein positive of the normal tunica mucosa esophageal tissues (26/38, 68.4%). There was statistical significance among the three groups (P<0.05). The sex, age, tumor size, or pathologic stage of the patients had no influence on the Syk protein (P>0.05).
2 By analysis of RT-PCR and Western-blot, Syk mRNA and protein expressed in TE-1,TE-13 and Eca-109 cells, and was strong positive in TE-1 cells,whereas not express in Yes-2 cells.
3 Through the detection of RT-PCR method, the expression of Syk gene was gradually decreased after treatment with Piceatannol of different concentrations (0、10、50μmol/L). As the Syk expression decreased, the ability of TE-1 cell migration and invasion was gradually recovered, and MMP-2, MMP-9 were up-regulated after treatment with Piceatannol of different concentrations.
4 The results of methylation-specific PCR showed that the promoter of Syk is hypermethylated in Yes-2 cells, while the hypermethylation cannot be detected in TE-1 cell.
Conclusions:
1 The mRNA and protein expression of Syk in ESCC were significantly higher than peri-cancer tissues and the normal tissues but not associate with the clinical pathology signs.
2 The down-regulated expression of Syk might be participate to the migration and invasion in TE-1 cells. The methylation of Syk promoter CpG island might be associated with the gene regulation of squamous cells.
方法:
1采用逆转录-聚合酶链反应(RT-PCR)检测30例食管鳞癌组织、27例食管鳞癌癌旁组织及30例正常食管黏膜组织中Syk mRNA的表达。免疫组化技术法检测39例食管鳞癌组织、27例食管鳞癌癌旁组织及38例正常食管黏膜组织中Syk蛋白的表达。分析Syk蛋白表达与临床病理学指标间的关系。
2用RT-PCR和Western-blot方法检测四种食管癌细胞株(Yes-2、Eca-109、TE-13和TE1)Syk的表达情况。
3给予不同浓度(0、10、50μmol/L)的Syk选择性抑制剂Piceatannol处理Syk阳性表达的TE-1细胞,观察Syk基因的表达变化;通过细胞划痕实验观察处理后细胞迁移能力的改变;采用RT-PCR法分析MMP-2、MMP-9的表达变化。
4 Syk基因甲基化状态检测:提取TE-1和Yes-2细胞DNA,经亚硫酸盐修饰纯化后,行甲基化特异性PCR(methylation-specific PCR, MSP)检测Syk启动子区的甲基化情况。
结果:
1 RT-PCR检测结果显示,在30例食管鳞癌、27例食管鳞癌癌旁组织和30例正常食管黏膜组织中,食管癌组织Syk mRNA表达量高于癌旁和正常黏膜组织,统计学分析表明,三组间Syk mRNA的表达差异无统计学意义。经抗Syk单抗进行免疫组化染色,Syk蛋白阳性反应为棕黄色颗粒,定位于细胞浆。结果显示在食管鳞癌组织中34例Syk蛋白表达阳性(34/39, 87.2%),癌旁组织18例表达阳性(18/27, 66.7%),正常食管黏膜组织中26例表达阳性(26/38, 68.4%),食管癌组织Syk蛋白表达高于食管癌旁组织和正常食管黏膜组织,三组间Syk蛋白表达差异具有统计学意义(P<0.05)。Syk蛋白表达与食管鳞癌患者年龄、性别、肿瘤大小、临床分期差异均无统计学意义(P均>0.05)。
2用RT-PCR和Western-blot方法检测四种不同食管癌细胞株,结果显示Syk基因和蛋白在食管癌细胞株TE1细胞中表达阳性,在Yes-2细胞中表达阴性,Eca-109、TE-13细胞表达为弱阳性。
3 Syk表达阳性的TE-1细胞在给予不同浓度(0、10、50μmol/L)Syk选择性抑制剂Piceatannol处理后,表达逐渐减弱;并且细胞迁移和侵袭能力逐渐恢复;MMP-2和MMP-9的表达逐渐上调。
4 MSP结果显示,TE-1细胞中未发现Syk基因启动子甲基化,Yes-2细胞中发现有Syk基因Syk基因启动子甲基化。
结论:
1 Syk基因和蛋白在食管鳞癌组织中表达增高,但Syk蛋白表达与临床病理学指标无关。
2 Syk表达下调可能参与了食管癌细胞的迁移、侵袭;且其启动子区cpG岛的甲基化参与了食管癌细胞的基因调控。
Objective: Cancer is a class of threatening diseases which has serious impact on health. Morbidity and mortality rate of malignant tumor is increasing in recent years. Tumorigenesis is a multifactor, multistep, polygene involved process that consists of complex pathological changes, such as activation of a variety of oncogenes, inactivation of tumor suppressor genes, and the disorder of other related regulatory factors. More than 50% of proto-oncogene and oncogene proteins have been characterized with the protein- tyrosine kinase (PTKs) activity. Spleen Tyrosine Kinase (Syk) is a non-receptor type of protein- tyrosine kinase that is widely expressed in hematopoietic cells, lymphocyte, fibroblast and vascular endothelial cells. Syk plays an essential role in lymphocyte development and activation. Recent studies have shown that Syk plays an important role in suppressing the growth and metastasis of human glandular epithelium malignant tumor cells with different origins. However, there was few studies paid attention to the expression of Syk in squamous cell carcinoma. In this study, we investigated the gene and protein expression of Syk in human esophageal squamous cell carcinoma (ESCC) and analyzed the effects of Syk on the biocharacteristics of ESCC.
Methods:
1 The expression of Syk gene in human esophageal carcinoma tissues (n=30), peri-cancer tissues (n=27) and normal tunica mucosa esophageal tissues (n=30) were detected by Reverse transcription-polymerase chain reaction (RT-PCR) technique. Immunohistochemistry was used to detect the expression of Syk protein in human esophageal carcinoma tissues (n=30), peri-cancer tissues (n=27) and normal tunica mucosa esophageal tissues (n=30). The relationships between the expression of Syk protein and clinical pathology features were analyzed.
2 RT-PCR and Western-blot was used to measure the expression of Syk gene and Syk protein in esophageal carcinoma cell lines of Yes-2, Eca-109, TE-13 and TE1 cells.
3 Expression of Syk gene was analyzed by RT-PCR, cell migrations were observed by wound healing assay, and expression of MMP-2, MMP-9 was detected by RT-PCR in TE-1 cells after treatment with Piceatannol of different concentrations (0、10、50μmol/L) .
4 The methylation status of the 5’CpG island located in the promoter region of Syk gene in esophageal carcinoma cell lines Yes-2 and TE-1 was detected by methylation specific PCR.
Results:
1 According to the results of RT-PCR, the mRNA expression of the Syk was higher in human esophageal carcinoma lesion tissues than the peri-cancer tissues and the normal esophageal tissues, but among the human esophageal carcinoma lesion tissues, peri-cancer tissues and normal tunica mucosa esophageal tissues the mRNA expression of the Syk had no statistical significance. By immunohistochemical staining with antibody of Sky, it was considered as positive if Syk was expressed in cytoplasm. 34 were Syk protein positive of the human esophageal carcinoma tissues samples (34/39, 87.2%), 18 Syk protein were detected in peri-cancer tissues (18/27, 66.7%) and 26 Syk protein positive of the normal tunica mucosa esophageal tissues (26/38, 68.4%). There was statistical significance among the three groups (P<0.05). The sex, age, tumor size, or pathologic stage of the patients had no influence on the Syk protein (P>0.05).
2 By analysis of RT-PCR and Western-blot, Syk mRNA and protein expressed in TE-1,TE-13 and Eca-109 cells, and was strong positive in TE-1 cells,whereas not express in Yes-2 cells.
3 Through the detection of RT-PCR method, the expression of Syk gene was gradually decreased after treatment with Piceatannol of different concentrations (0、10、50μmol/L). As the Syk expression decreased, the ability of TE-1 cell migration and invasion was gradually recovered, and MMP-2, MMP-9 were up-regulated after treatment with Piceatannol of different concentrations.
4 The results of methylation-specific PCR showed that the promoter of Syk is hypermethylated in Yes-2 cells, while the hypermethylation cannot be detected in TE-1 cell.
Conclusions:
1 The mRNA and protein expression of Syk in ESCC were significantly higher than peri-cancer tissues and the normal tissues but not associate with the clinical pathology signs.
2 The down-regulated expression of Syk might be participate to the migration and invasion in TE-1 cells. The methylation of Syk promoter CpG island might be associated with the gene regulation of squamous cells.
引文
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12 Tsuchida S, Yanagi S, Inatome R, et al. Purification of a 72-kDa protein- tyrosine kinase from rat liver and its identification as Syk: involvement of Syk in signaling events of hepatocytes. J Biochem (Tokyo), 2000, 127(2): 321~327
13 Hoeller C, Thallinger C, Pratschei B, et al. The non-receptor- associated tyrosine kinase Syk is a regulator of metastatic behavior in human melanoma cells. J Invest Dermatol, 2005, 124(6): 1293~1299
14陈明霞,丁永斌,蒋奎荣,等.酪氨酸激酶Syk在胰腺癌中的表达检测及其临床意义.南京医科大学学报(自然科学版), 2003, 23(5): 492~493
15 Moroni M, Soldatenkov V, Zhang L, et al. Progressive loss of Syk and abnormal proliferation in breast cancer cells. Cancer Res, 2004, 64 (20): 7346~7354
16 Thompson E W, Paik S, Brunner N, et al. Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J Cell Physiol, 1992, 150; 534~544
17 Nakashima H, Natsugoe S, Ishigami S, et al. Clinical significance of nuclear expression of spleen tyrosine kinase (Syk) in gastric cancer. Cancer Lett, 2006, 236 (1): 89~94
18 Wang S, Ding YB, Chen GY, et al. Hypermethylation of Syk gene in romoter region associated with oncogenesis and metastasis of gastric carcino- ma. World J Gast roenterol, 2004, 10(12): 1815~1818
19夏建国,丁永斌,陈国玉.胃癌候选抑癌基因Syk表达的意义.世界华人消化杂志, 2004, 12(4): 767~769
20 Murphy MA, Schnall RG, Venter DJ, et al. Tissue hyperplasia and enhanced T cell signaling via ZAP-70 in c-Cbl-deficient mice. Mol Cell Biol, 1998, 18(8): 4872~4882
21 Tracy Layton, Cristel Stalens, Felizza Gunderson, et al. Syk Tyrosine Kinase Acts as a Pancreatic Adenocarcinoma Tumor Suppressor by Regulating Cellular Growth and Invasion. Am J Pathol, 2009, 175(6): 2625~2636
22宋玉琴,周武元,赵文华,等. Syk蛋白在胰腺癌组织中的表达及临床意义.中华肿瘤防治杂志, 2007, 14(15): 1149~1151
23万伟,吴小鹏,王建平,等. Syk基因的表达与大肠癌关系的研究.中国现代普通外科进展, 2006, 9(l): 40~42
24 Sutima L, Carol B, Lisa P,et al. Syk tyrosine kinase is linked to cell motility and progression in squamous cell carcinomas of the head and neck. Cancer Res, 2007, 67 (16): 7907~7916.
25刘泓基,臧真真,侯巧燕,等. Syk基因在食管鳞癌组织中的表达及临床意义.世界华人消化杂志, 2010, 18(1): 33~38
26 Slack A, Cervoni N, Pinard M, et al. Feedback regulation of DNA methyltransferase gene expression by methylation. Eur J Biochem, 1999, 264: 191~199
27 McCluskey LL, Chen C, Delgadillo E, et al. Differences in p16 gene methylation and expression in benign and malignant ovarian tumors. Gynecol Oncol, 1999 (72): 87~92
28 Yuan Y, Mendez R, Sahin A, et al. Hypermethylation leads to silencing of the SYK gene in human breast cancer. Cancer Res, 2001, 61(14): 5558~ 5561
29 Ding YB, Wu ZY, Wang S, et al. Hypermethylation of syk gene in promoter region is associated with oncogenesis, metastasis of breast carcinoma. Natl Med J China, 2004, 84(4): 290~293
30 Wang S, Ding YB, Chen GY, et al. Hypermethylation of Syk gene inpromoter region associated with oncogenesis and metastasis of gastric carcinoma. World J Gastroenterol, 2004, 10(12):1815~1818
31杨祖立,汪建平,兀云飞,等.结直肠侵袭性与脾酪氨酸激酶基因Syk启动子甲基化的关系.中华实验外科杂志, 2005, 22(1): 68~70
32 Wallner M, Herbst A, Behrens A, et al. Methylation of Serum DNA Is an Independent Prognostic Marker in Colorectal Cancer. Clin Cancer Res, 2006, 12(24): 7347~7355
33 Yuan Y, Wang J, Li J, et al. Frequent epigenetic inactivation of spleen tyrosine kinasc gene in human hepatocellular carcinoma. Clin Cancer Res, 2006, 12(22): 6687~6695
34 Goodman PA, Burkhardt N, Juran B, et al. Hypermethylation of the spleen tyrosine kinase promoter in T-lineage acute lymphoblastic leukemia. Oncogene, 2003, 22(16): 2504~2514
35 Ji R C. Lymphatic endothelial cells, lymphangiogenesis, end extracellular matrix. Lymphat Res Biol, 2006, 4 (2): 83~100
36 Abtahian F, Guerriero A, Sebzda E, et al. Regulation of blood and lymphatic vascular separation by signaling proteins SLP-76 and Syk. Science, 2003, 299(5604): 247~251
37 Carter W B, Hoyiug J B, Boswell C, et al. HER2/neu over-expression induces endothelial cell retraction. Int J Cancer, 2001, 91(3): 295~299
38 Mahabeleshwar GH, Kundu GC. Syk, a protein-tyrosine kinase suppresses the cell motility and nuclear factor kappa B-mediated secretion of urokinase type plas minogen activator by inhibiting the phosphatidy- linositol 3'-kinase activity in breast cancer cells. J Biol Chem, 2003, 278 (8): 6209~6221
39 Hoeller C, Thallinger C, Pratscher B, et al. The non-receptor- associated tyrosine kinase Syk is a regulator of metastatic behavior in human melanoma cells. J Invest Dermatol, 2005, 124(6): 1293~1299
2 Coopman PJ, Do MT, Barth M, et al. The Syk tyrosine kinase suppresses malignant growth of human breast cancer cells. Nature, 2000, 406(6797): 742~747
3 Toyama T, Iwase H, Yamashita H, et al. Reduced expression of the Syk gene is correlated with poor prognosis in human breast cancer. Cancer Lett, 2003, 189(1): 97~102
4 Moroni M, Soldatenkov V, Zhang L, et al. Progressive loss of Syk and abnormal proliferation in breast cancer cells. Cancer Res, 2004, 64 (20): 7346~7354
5 Nakashima H, Natsugoe S, Ishigami S, et al. Clinical significance of nuclear expression of spleen tyrosine kinase (Syk) in gastric cancer. Cancer Lett, 2006, 236 (1): 89~94
6袁世龙,郭春良,焦辉,等. Syk与胃癌生物学行为的相关性研究.武警医学院学报, 2007, 5(16): 514~517
7 Layton T, Stalens C, Gunderson F, et al. Syk tyrosine kinase acts as a pancreatic adenocarcinoma tumor suppressor by regulating cellular growthand invasion. Am J Pathol , 2009, 175(6): 2625~2636
8陈明霞,丁永斌,蒋奎荣,等.酪氨酸激酶Syk在胰腺癌中的表达检测及其临床意义.南京医科大学学报(自然科学版), 2003, 23(5): 492~493
9宋玉琴,周武元,赵文华,等. Syk蛋白在胰腺癌组织中的表达及临床意义.中华肿瘤防治杂志, 2007, 14(15): 1149~1151
10万伟,吴小鹏,王建平,等. Syk基因的表达与大肠癌关系的研究.中国现代普通外科进展, 2006, 9(l): 40~42
11 Taniguchi T, Kobayashi T, Kondo J, et al. Molecular cloning of a porcine gene Syk that encodes a 72-kDa protein-tyrosine kinase showing high susceptibility to proteolysis. J Biol Chem, 1991, 266(24): 15790~15796
12齐义新,胡洁,宋振川,等.乳腺癌组织中Syk表达及其与临床病理关系研究.中国肿瘤临床, 2005, 32(24): 1386~1388
13 Sutima L, Carol B, Lisa P, et al. Syk tyrosine kinase is linked to cell motility and progression in squamous cell carcinomas of the head and neck. Cancer Res, 2007, 67(16): 7907~7916
14 Wang L, Duke L, Zhang PS, et al. Alternative splicing disrupts a nuclear localization signal in spleen tyrosine kinase that is required for invasion suppression in breast cancer. Cancer Res, 2003, 63(15): 4724~4730
15 Wang L, Devarajan E, He J, et al. Transcription repressor activity of spleen tyrosine kinase mediates breast tumor suppression. Cancer Res, 2005, 65(22): 10289~10297
16刘泓基,臧真真,侯巧燕,等. Syk基因在食管鳞癌组织中的表达及临床意义.世界华人消化杂志, 2010, 18(1): 33~38
1 Coopman PJ, Do MT, Barth M, et al. The Syk tyrosine kinase suppresses malignant growth of human breast cancer cells. Nature, 2000, 406 (6797): 742~747
2 Lu J, Lin WH, Chen SY, et al. Syk tyrosine kinase mediates Epstein-Barr virus latent membrane protein 2A-induced cell migration in epithelial cells. J Biol Chem , 2006, 281: 8806~8814
3 Carter W B, Hoyiug J B, Boswell C, et al. HER2/neu over-expression induces endothelial cell retraction. Int J Cancer, 2001, 91(3): 295~299
4 Freya W, Antje C, Bora A, et al. Piceatannol, a Natural Analog of Resveratrol, Inhibits Progression through the S Phase of the Cell Cycle in Colorectal Cancer Cell Lines. J Nutr, 2002, 132: 298~302
5 Ashikawa K, Majumdar S, Banerjee S, et al. Piceatannol inhibits TNF-induced NF-κB activation and NF-κB-mediated gene expression through suppression of InBa kinase and p65 phosphorylation. Immunol, 2002, 169:6490~6497
6 Dang O, Navarro L, David M. Inhibition of lipopolysaccharide-induced interferon regulatory factor 3 activation and protection from septic shock by hydroxystilbenes. Shock, 2004, 21:470~475
7 Matsuda H, Tewtrakul S, Morikawa T, et al. Anti-allergic activity of stilbenes from Korean rhubarb (Rheum undulatum L): structure require- ments for inhibition of antigen-induced degranulation and their effects on the release of TNF-αand IL-4 in RBL-2H3 cells. Bioorg Med Chem, 2004,12: 4871~4876
8 Seow CJ, Chue SC, Duan W, et al. Effects of inhibitors of the tyrosine signaling cascade on antigen challenge of guinea pig airways in vitro. Ann Acad Med Singapore, 2004, 33: 41~43
9 Wieder T, Prokop A, Bagci B, et al. Piceatannol, a hydroxylated analog of the chemopreventive agent resveratrol, is a potent inducer of apoptosis in the lymphoma cell line BJAB and in primary, leukemic lymphoblasts. Leukemia, 2001, 15: 1735~1742
10 Larrosa M, Tomas-Barberan FA, Espin JC. The grape and wine poly- phenol piceatannol is a potent inducer of apoptosis in human SK-Mel-28 melanoma cells. Eur J Nutr, 2004, 43: 275~284
11 Barton BE, Karras JG, Murphy TF, et al. Signal transducer and activator of transcription3 (STAT3) activation in prostate cancer: direct STAT3 inhibition induces apoptosis in prostate cancer lines. Mol Cancer Ther, 2004, 3: 11~20
12 Ferrigni NR, McLaughlin JL, Powell RG, et al. Use of potato disc and brine shrimp bioassays to detect activity and isolate piceatannol as the antileukemic principle from the seeds of Euphorbia lagascae. Nat Prod, 1984, 47: 347~52
13 Geahlen RL, McLaughlin JL, et al. Piceatannol (3,4,3’,5’-tetrahydroxy- transstilbene) is a naturally occurring protein-tyrosine kinase inhibitor. Biochem Biophys Res Commun, 1989, 165:241~245
14 Thakkar K, Geahlen RL, Cushman M. Synthesis and protein-tyrosine kinase inhibitory activity of polyhydroxylated stilbene analogues of piceatannol. J Med Chem, 1993, 36: 2950~2955
15 Law DA, Nannizzi-Alaimo L, Ministri K, et al. Genetic and pharma- cological analyses of Syk function inαIIbβ3 signaling in platelets. Blood, 1999, 93: 2645~2652
16 Wang B H, Lu Z X, Polya G M. Inhibition of eukaryote serine/threonine- specific protein kinases by piceatannol. Planta Med, 1998, 64: 195~199.
17 Zheng J, Ramirez V D. Piceatannol, a stilbene phytochemical, inhibitsmitochondrial F0F1-ATPase activity by targeting the F1 complex. Biochem. Biophys. Res. Commun, 1999, 261: 499~503
18 Mahabeleshwar G H, Kundu G C. Syk, a protein-tyrosine kinase, suppresses the cell motility and nuclear factor kappa B-mediated secretion of urokinase type plasminogen activator by inhibiting the phosphatidy- linositol 3'-kinase activity in breast cancer cells. J Biol Chem, 2003, 278 (8): 6209~6221
19 L Wang, L Duke, P S Zhang, et al. Alternative splicing disruptsa nuclear localization signal in spleen tyrosine kinase that isrequired for invasion suppression in breast cancer. Cancer Res, 2003, 63: 4724~4730
20 Sutima Luangdilok, Carol Box, Lisa Patterson, et al. Syk Tyrosine Kinase Is Linked to Cell Motility and Progression in Squamous Cell Carcinomas of the Head and Neck. Cancer Res, 2007; 67: 7907~7916
1 Martin F, Gisela Z. Anne-Catherine Andres and Andrew Ziemiecki: Molecule characterization of the murine SYK protein tyrosine kinase cDNA, transcripts and protein. Biochemical and biophysical research communications, 1995, 213: 273~281
2 Taniguchi T, Kobayashi T, Kondo J, et al. Molecular cloning of a porcine gene Syk that encodes a 72-kDa protein-tyrosine kinase showing high susceptibility to proteolysis. J Biol Chem, 1991, 266(24): 15790~15796
3 Turner M, Mee P J and Costello P S. Perinatal lethality and blocked B cell development in mice lacking the tyrosine kinase Syk. Nature, 1995, 378: 298~302
4 Wang L, Duke L, Zhang PS, et al. Alternative splicing disrupts a nuclear localization signal in spleen tyrosine kinase that is required for invasion suppression in breast cancer. Cancer Res, 2003, 63(20): 4724~4730
5 Turner M, Schweighoffer E, Colucei F, et al. Tyrosine kinase SYK, essential functions for immunoreceptor signaling. Immunol Today, 2000, 21: 148~154
6 Sada K, Takano T, Yanagi S, et al. Structure and function of Syk protein- tyrosine kinase. J Biochem, 2001, 130 (2): 177~186
7 Coopman PJ, Do MT, Barth M, et al. The Syk tyrosine kinase suppresses malignant growth of human breast cancer cells. Nature, 2000, 406 (6797): 742~747
8 Ulanova M, Puttagunta L, Marcet-Palacios M, et al. Syk tyrosine kinase participates in betal-integrin signaling and inflammatory responses in airway epithelial cells.Am J Physiol Lung Cell Mol Physiol, 2005, 288(3): 497~507
9 Yamada T, Fujieda S, Yanagi S, et al. Protein-tyrosine kinase Syk expressed in human nasal frbroblasts and its effect on RANTES production . Immunol, 2001, 166(1): 538~543
10 Inatome R, Yanagi S, Takano T, et al. A critical role for Syk in endothelial cell proliferation and migration. Biochem Biophys Res Commun, 2001, 286(1): 195~199
11 Tsujimura T, Yanagi S, Inatome R, et al. Syk protein-tyrosine kinase is involved in neuron-like differentiation of embryonal carcinoma P19 cells. FEBS Lett, 2001, 489: 129~133
12 Tsuchida S, Yanagi S, Inatome R, et al. Purification of a 72-kDa protein- tyrosine kinase from rat liver and its identification as Syk: involvement of Syk in signaling events of hepatocytes. J Biochem (Tokyo), 2000, 127(2): 321~327
13 Hoeller C, Thallinger C, Pratschei B, et al. The non-receptor- associated tyrosine kinase Syk is a regulator of metastatic behavior in human melanoma cells. J Invest Dermatol, 2005, 124(6): 1293~1299
14陈明霞,丁永斌,蒋奎荣,等.酪氨酸激酶Syk在胰腺癌中的表达检测及其临床意义.南京医科大学学报(自然科学版), 2003, 23(5): 492~493
15 Moroni M, Soldatenkov V, Zhang L, et al. Progressive loss of Syk and abnormal proliferation in breast cancer cells. Cancer Res, 2004, 64 (20): 7346~7354
16 Thompson E W, Paik S, Brunner N, et al. Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J Cell Physiol, 1992, 150; 534~544
17 Nakashima H, Natsugoe S, Ishigami S, et al. Clinical significance of nuclear expression of spleen tyrosine kinase (Syk) in gastric cancer. Cancer Lett, 2006, 236 (1): 89~94
18 Wang S, Ding YB, Chen GY, et al. Hypermethylation of Syk gene in romoter region associated with oncogenesis and metastasis of gastric carcino- ma. World J Gast roenterol, 2004, 10(12): 1815~1818
19夏建国,丁永斌,陈国玉.胃癌候选抑癌基因Syk表达的意义.世界华人消化杂志, 2004, 12(4): 767~769
20 Murphy MA, Schnall RG, Venter DJ, et al. Tissue hyperplasia and enhanced T cell signaling via ZAP-70 in c-Cbl-deficient mice. Mol Cell Biol, 1998, 18(8): 4872~4882
21 Tracy Layton, Cristel Stalens, Felizza Gunderson, et al. Syk Tyrosine Kinase Acts as a Pancreatic Adenocarcinoma Tumor Suppressor by Regulating Cellular Growth and Invasion. Am J Pathol, 2009, 175(6): 2625~2636
22宋玉琴,周武元,赵文华,等. Syk蛋白在胰腺癌组织中的表达及临床意义.中华肿瘤防治杂志, 2007, 14(15): 1149~1151
23万伟,吴小鹏,王建平,等. Syk基因的表达与大肠癌关系的研究.中国现代普通外科进展, 2006, 9(l): 40~42
24 Sutima L, Carol B, Lisa P,et al. Syk tyrosine kinase is linked to cell motility and progression in squamous cell carcinomas of the head and neck. Cancer Res, 2007, 67 (16): 7907~7916.
25刘泓基,臧真真,侯巧燕,等. Syk基因在食管鳞癌组织中的表达及临床意义.世界华人消化杂志, 2010, 18(1): 33~38
26 Slack A, Cervoni N, Pinard M, et al. Feedback regulation of DNA methyltransferase gene expression by methylation. Eur J Biochem, 1999, 264: 191~199
27 McCluskey LL, Chen C, Delgadillo E, et al. Differences in p16 gene methylation and expression in benign and malignant ovarian tumors. Gynecol Oncol, 1999 (72): 87~92
28 Yuan Y, Mendez R, Sahin A, et al. Hypermethylation leads to silencing of the SYK gene in human breast cancer. Cancer Res, 2001, 61(14): 5558~ 5561
29 Ding YB, Wu ZY, Wang S, et al. Hypermethylation of syk gene in promoter region is associated with oncogenesis, metastasis of breast carcinoma. Natl Med J China, 2004, 84(4): 290~293
30 Wang S, Ding YB, Chen GY, et al. Hypermethylation of Syk gene inpromoter region associated with oncogenesis and metastasis of gastric carcinoma. World J Gastroenterol, 2004, 10(12):1815~1818
31杨祖立,汪建平,兀云飞,等.结直肠侵袭性与脾酪氨酸激酶基因Syk启动子甲基化的关系.中华实验外科杂志, 2005, 22(1): 68~70
32 Wallner M, Herbst A, Behrens A, et al. Methylation of Serum DNA Is an Independent Prognostic Marker in Colorectal Cancer. Clin Cancer Res, 2006, 12(24): 7347~7355
33 Yuan Y, Wang J, Li J, et al. Frequent epigenetic inactivation of spleen tyrosine kinasc gene in human hepatocellular carcinoma. Clin Cancer Res, 2006, 12(22): 6687~6695
34 Goodman PA, Burkhardt N, Juran B, et al. Hypermethylation of the spleen tyrosine kinase promoter in T-lineage acute lymphoblastic leukemia. Oncogene, 2003, 22(16): 2504~2514
35 Ji R C. Lymphatic endothelial cells, lymphangiogenesis, end extracellular matrix. Lymphat Res Biol, 2006, 4 (2): 83~100
36 Abtahian F, Guerriero A, Sebzda E, et al. Regulation of blood and lymphatic vascular separation by signaling proteins SLP-76 and Syk. Science, 2003, 299(5604): 247~251
37 Carter W B, Hoyiug J B, Boswell C, et al. HER2/neu over-expression induces endothelial cell retraction. Int J Cancer, 2001, 91(3): 295~299
38 Mahabeleshwar GH, Kundu GC. Syk, a protein-tyrosine kinase suppresses the cell motility and nuclear factor kappa B-mediated secretion of urokinase type plas minogen activator by inhibiting the phosphatidy- linositol 3'-kinase activity in breast cancer cells. J Biol Chem, 2003, 278 (8): 6209~6221
39 Hoeller C, Thallinger C, Pratscher B, et al. The non-receptor- associated tyrosine kinase Syk is a regulator of metastatic behavior in human melanoma cells. J Invest Dermatol, 2005, 124(6): 1293~1299