NDRG2截短肽的抗肿瘤作用及NDRG2与胶质瘤预后的关系
|
摘要
中枢神经系统肿瘤,尤其是胶质瘤,以其恶劣的预后严重地危险着人们的健康。寻求疗效更佳的治疗方案和更可靠的预后评估因子,是神经科学家和临床医生们一直的追求。
在这个过程中,我校研究人员首先发现了人类的NDRG2基因。该基因在包括胶质瘤在内的多种肿瘤中低表达,其表达受到经典癌基因MYC的负调控;人为调高NDRG2在肿瘤细胞中的表达,会对肿瘤细胞的增殖产生明显的抑制作用;基于上述发现,NDRG2被认为是一种抑癌候选基因,受到我们的持续关注。
一方面,通过对NDRG2的解构分析,我们希望发现其作用的具体机制;另一方面,我们也想知道该基因的表达状态对于肿瘤患者的实际意义如何。
本项研究的第一部分是从人类NDRG2蛋白的分子分析入手,预测其三维结构为两个结构域构成的球性蛋白,根据结构域,设计出若干分别代表其N末端、C末端,以及两个结构域的截短肽片段,构建其真核表达载体,转染胶质瘤细胞系U87MG和U251以及经典肿瘤细胞HeLa,通过流式细胞术观察肿瘤细胞的凋亡、周期阻滞和增殖(MTT法,仅对HeLa)情况,发现C末端的129肽对NDRG2发挥抗肿瘤作用最为关键,且N、C末端折叠成的结构域和C末端自身都可在某些情况下发挥较全长蛋白更强的抗肿瘤作用,且以诱导细胞周期阻滞为主。
在本项研究的第二部分,我们以一个时间段于我院进行常规手术治疗和放、化疗的胶质瘤患者为研究对象,通过免疫组化分析其肿瘤组织中的NDRG2的表达情况,比较其表达与肿瘤分级的关系;并通过对NDRG2表达和术后患者生存状态、生活质量的相关分析,对NDRG2作为预后因子的可能性进行评价,结果表明:NDRG2蛋白表达与病理分级呈负相关;与星形细胞瘤患者的术后累计生存率、生存时间、生活质量呈正相关,可以作为独立危险因素来评估胶质瘤患者的预后。
总之,通过这项研究,我们对NDRG2发挥作用的关键部位进行了定位,也评价了NDRG2在胶质瘤预后判断中的价值,既为今后机制研究明确了方向,也为其作为预后指标及治疗手段而应用于临床奠定了基础。
The awful prognosis of glioma is serious threaten for people’s heath. It is of a persistent work for all neuroscientists and neurologists to pursue the reliable prognositic biomarker and the new therapies with well-prognosis.
Our team members found the human NDRG2 gene first and confirmed its low expression in many tumor tissues and cell lines. It is downstream regulated by MYC, and inhibits the cell proliferation when it is transfected into glioma cell lines and induced to overexpress. Thus the gene is regarded as a candidate for tumor suppressor gene and attracts our attention for a long time.
Two questions focus our attention on the present study: Which part of the NDRG2 plays the leading role in its antitumor function? What correlation exists between the expression of NDRG2 and the patients’overall survival?
The first part of the present study is about the correlation between the structure of NDRG2 and its antitumor function. Here, we started our research from the analysis for the molecular structure of NDRG2, predicted its 3-dimentional structure according to its homologous protein of mouse. Then, we truncated the NDRG2 protein into several parts theoretically, designed and constructed the eukaryotic expression plasimds for each truncated peptides of NDRG2. Two glioma cell lines (U87MG and U251) and a classical cervical cancer cell line (HeLa) were transfected by the plasmids and their cell cycle arrest and apoptosis were dectected by flowcytometry. More than that, the proliferation of HeLa cells after being transfected by our plasminds was also observed by MTT assay. We found that the C-terminal of NDRG2 plays the leading role in its antitumor fuction, and sometimes the truncated Peptide AC and Peptide C showed more powerful antitumor function than the whole length protein. Peptide AC is the mimic of the big domain of original NDRG2 protein and Peptide C is consisted of 129 amino acids adjacent to C-terminal.
In the second part, we studied the expression profile of NDRG2 in human astrocytoma tissues by immunohistochemistry and found it was significantly downregulated in astrocytomas. The level of expression of NDRG2 showed an inverse correlation with the WHO grade of tumors and a positive one with the life span of patients. Our results suggested that NDRG2 may play a pivotal role as a potent tumor suppressor and a prognostic biomarker candidate for human astrocytoma.
In brief, we located the part playing the leading role for NDRG2 antitumor function and assessed the value of NDRG2 as prognostic biomarker in the study.
在这个过程中,我校研究人员首先发现了人类的NDRG2基因。该基因在包括胶质瘤在内的多种肿瘤中低表达,其表达受到经典癌基因MYC的负调控;人为调高NDRG2在肿瘤细胞中的表达,会对肿瘤细胞的增殖产生明显的抑制作用;基于上述发现,NDRG2被认为是一种抑癌候选基因,受到我们的持续关注。
一方面,通过对NDRG2的解构分析,我们希望发现其作用的具体机制;另一方面,我们也想知道该基因的表达状态对于肿瘤患者的实际意义如何。
本项研究的第一部分是从人类NDRG2蛋白的分子分析入手,预测其三维结构为两个结构域构成的球性蛋白,根据结构域,设计出若干分别代表其N末端、C末端,以及两个结构域的截短肽片段,构建其真核表达载体,转染胶质瘤细胞系U87MG和U251以及经典肿瘤细胞HeLa,通过流式细胞术观察肿瘤细胞的凋亡、周期阻滞和增殖(MTT法,仅对HeLa)情况,发现C末端的129肽对NDRG2发挥抗肿瘤作用最为关键,且N、C末端折叠成的结构域和C末端自身都可在某些情况下发挥较全长蛋白更强的抗肿瘤作用,且以诱导细胞周期阻滞为主。
在本项研究的第二部分,我们以一个时间段于我院进行常规手术治疗和放、化疗的胶质瘤患者为研究对象,通过免疫组化分析其肿瘤组织中的NDRG2的表达情况,比较其表达与肿瘤分级的关系;并通过对NDRG2表达和术后患者生存状态、生活质量的相关分析,对NDRG2作为预后因子的可能性进行评价,结果表明:NDRG2蛋白表达与病理分级呈负相关;与星形细胞瘤患者的术后累计生存率、生存时间、生活质量呈正相关,可以作为独立危险因素来评估胶质瘤患者的预后。
总之,通过这项研究,我们对NDRG2发挥作用的关键部位进行了定位,也评价了NDRG2在胶质瘤预后判断中的价值,既为今后机制研究明确了方向,也为其作为预后指标及治疗手段而应用于临床奠定了基础。
The awful prognosis of glioma is serious threaten for people’s heath. It is of a persistent work for all neuroscientists and neurologists to pursue the reliable prognositic biomarker and the new therapies with well-prognosis.
Our team members found the human NDRG2 gene first and confirmed its low expression in many tumor tissues and cell lines. It is downstream regulated by MYC, and inhibits the cell proliferation when it is transfected into glioma cell lines and induced to overexpress. Thus the gene is regarded as a candidate for tumor suppressor gene and attracts our attention for a long time.
Two questions focus our attention on the present study: Which part of the NDRG2 plays the leading role in its antitumor function? What correlation exists between the expression of NDRG2 and the patients’overall survival?
The first part of the present study is about the correlation between the structure of NDRG2 and its antitumor function. Here, we started our research from the analysis for the molecular structure of NDRG2, predicted its 3-dimentional structure according to its homologous protein of mouse. Then, we truncated the NDRG2 protein into several parts theoretically, designed and constructed the eukaryotic expression plasimds for each truncated peptides of NDRG2. Two glioma cell lines (U87MG and U251) and a classical cervical cancer cell line (HeLa) were transfected by the plasmids and their cell cycle arrest and apoptosis were dectected by flowcytometry. More than that, the proliferation of HeLa cells after being transfected by our plasminds was also observed by MTT assay. We found that the C-terminal of NDRG2 plays the leading role in its antitumor fuction, and sometimes the truncated Peptide AC and Peptide C showed more powerful antitumor function than the whole length protein. Peptide AC is the mimic of the big domain of original NDRG2 protein and Peptide C is consisted of 129 amino acids adjacent to C-terminal.
In the second part, we studied the expression profile of NDRG2 in human astrocytoma tissues by immunohistochemistry and found it was significantly downregulated in astrocytomas. The level of expression of NDRG2 showed an inverse correlation with the WHO grade of tumors and a positive one with the life span of patients. Our results suggested that NDRG2 may play a pivotal role as a potent tumor suppressor and a prognostic biomarker candidate for human astrocytoma.
In brief, we located the part playing the leading role for NDRG2 antitumor function and assessed the value of NDRG2 as prognostic biomarker in the study.
引文
1. Shimono A, Okuda T, Kondoh H. N-myc-dependent repression of ndr1, a gene identified by direct subtraction of whole mouse embryo cDNAs between wild type and N-myc mutant. Mech Dev, 1999, 83(1-2): 39～52
2. Okuda T, Kondoh H. Identification of new genes ndr2 and ndr3 which are related to Ndr1/RTP/Drg1 but show distinct tissue specificity and response to N-myc. Biochem Biophys Res Commun, 1999, 266(1): 208～215
3. Kokame K, Kato H, Miyata T. Homocysteine-respondent genes in vascular endothelial cells identified by differential display analysis. GRP78/BiP and novel genes. J Biol Chem, 1996, 271(47): 29659～29665
4. van Belzen N, Dinjens W N, Diesveld M P, et al. A novel gene which is up-regulated during colon epithelial cell differentiation and down-regulated in colorectal neoplasms. Lab Invest, 1997, 77(1): 85～92
5. Lin T M, Chang C. Cloning and characterization of TDD5, an androgen target gene that is differentially repressed by testosterone and dihydrotestosterone. Proc Natl Acad Sci U S A, 1997, 94(10): 4988～4993
6. Zhou D, Salnikow K, Costa M. Cap43, a novel gene specifically induced by Ni2+ compounds. Cancer Res, 1998, 58(10): 2182～2189
7.邓艳春,药立波,刘新平, et al.人脑内-含有ACP样结构域新基因的发现.生物化学与生物物理进展, 2001, 28(1): 72～76
8. Nagase T, Ishikawa K, Suyama M, et al. Prediction of the coding sequences of unidentified human genes. XIII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res, 1999, 6(1): 63～70
9. Zhao W, Tang R, Huang Y, et al. Cloning and expression pattern of thehuman NDRG3 gene. Biochim Biophys Acta, 2001, 1519(1-2): 134～138
10. Zhou R H, Kokame K, Tsukamoto Y, et al. Characterization of the human NDRG gene family: a newly identified member, NDRG4, is specifically expressed in brain and heart. Genomics, 2001, 73(1): 86～97
11. Qu X, Jia H, Garrity D M, et al. Ndrg4 is required for normal myocyte proliferation during early cardiac development in zebrafish. Dev Biol, 2008, 317(2): 486～496
12. Krauter-Canham R, Bronner R, Evrard J, et al. A transmitting tissue-and pollen-expressed protein from sunflower with sequence similarity to the human RTP protein. Plant Sci, 1997, 129: 191～202
13. Qu X, Zhai Y, Wei H, et al. Characterization and expression of three novel differentiation-related genes belong to the human NDRG gene family. Mol Cell Biochem, 2002, 229(1-2): 35～44
14. Shaw E, Mccue L A, Lawrence C E, et al. Identification of a novel class in the alpha/beta hydrolase fold superfamily: the N-myc differentiation-related proteins. Proteins, 2002, 47(2): 163～168
15. Okuda T, Kokame K, Miyata T. Differential expression patterns of NDRG family proteins in the central nervous system. J Histochem Cytochem, 2008, 56(2): 175～182
16. Hou W G, Zhao Y, Shen L, et al. Differential expression of N-Myc downstream regulated gene 2 (NDRG2) in the rat testis during postnatal development. Cell Tissue Res, 2009, 337(2): 257～267
17. Hou W, Hu J, Li Y, et al. Altered Expression of NDRG2 in the Testes of Experimental Rat Model of Cryptorchidism. Urology, 2010,
18. Lachat P, Shaw P, Gebhard S, et al. Expression of NDRG1, a differentiation-related gene, in human tissues. Histochem Cell Biol, 2002,118(5): 399～408
19. Wang W, Li Y, Li Y, et al. NDRG3 is an androgen regulated and prostate enriched gene that promotes in vitro and in vivo prostate cancer cell growth. Int J Cancer, 2009, 124(3): 521～530
20. Deng Y, Yao L, Chau L, et al. N-Myc downstream-regulated gene 2 (NDRG2) inhibits glioblastoma cell proliferation. Int J Cancer, 2003, 106(3): 342～347
21.胡晓兰,药立波,张远强, et al. NDRG2在人胚胎组织中的表达分布特点.生理学报, 2006, 58(04): 331～336
22. Shen L, Zhao Z Y, Wang Y Z, et al. Immunohistochemical detection of Ndrg2 in the mouse nervous system. Neuroreport, 2008, 19(9): 927～931
23. Hu X L, Liu X P, Deng Y C, et al. Expression analysis of the NDRG2 gene in mouse embryonic and adult tissues. Cell Tissue Res, 2006, 325(1): 67～76
24. Wang L, Liu N, Yao L, et al. NDRG2 is a new HIF-1 target gene necessary for hypoxia-induced apoptosis in A549 cells. Cell Physiol Biochem, 2008, 21(1-3): 239～250
25. Sugiki T, Murakami M, Taketomi Y, et al. N-myc downregulated gene 1 is a phosphorylated protein in mast cells. Biol Pharm Bull, 2004, 27(5): 624～627
26. Unoki M, Nakamura Y. Growth-suppressive effects of BPOZ and EGR2, two genes involved in the PTEN signaling pathway. Oncogene, 2001, 20(33): 4457～4465
27. Guan R J, Ford H L, Fu Y, et al. Drg-1 as a differentiation-related, putative metastatic suppressor gene in human colon cancer. Cancer Res, 2000, 60(3): 749～755
28. Bandyopadhyay S, Pai S K, Gross S C, et al. The Drg-1 gene suppresses tumor metastasis in prostate cancer. Cancer Res, 2003, 63(8): 1731～1736
29. Bandyopadhyay S, Pai S K, Hirota S, et al. Role of the putative tumor metastasis suppressor gene Drg-1 in breast cancer progression. Oncogene, 2004, 23(33): 5675～5681
30. Bandyopadhyay S, Wang Y, Zhan R, et al. The tumor metastasis suppressor gene Drg-1 down-regulates the expression of activating transcription factor 3 in prostate cancer. Cancer Res, 2006, 66(24): 11983～11990
31. Kovacevic Z, Richardson D R. The metastasis suppressor, Ndrg-1: a new ally in the fight against cancer. Carcinogenesis, 2006, 27(12): 2355～2366
32. Ellen T P, Ke Q, Zhang P, et al. NDRG1, a growth and cancer related gene: regulation of gene expression and function in normal and disease states. Carcinogenesis, 2008, 29(1): 2～8
33. Piquemal D, Joulia D, Balaguer P, et al. Differential expression of the RTP/Drg1/Ndr1 gene product in proliferating and growth arrested cells. Biochim Biophys Acta, 1999, 1450(3): 364～373
34. Angst E, Sibold S, Tiffon C, et al. Cellular differentiation determines the expression of the hypoxia-inducible protein NDRG1 in pancreatic cancer. Br J Cancer, 2006, 95(3): 307～313
35. Gomez-Casero E, Navarro M, Rodriguez-Puebla M L, et al. Regulation of the differentiation-related gene Drg-1 during mouse skin carcinogenesis. Mol Carcinog, 2001, 32(2): 100～109
36. Tschan M P, Shan D, Laedrach J, et al. NDRG1/2 expression is inhibited in primary acute myeloid leukemia. Leuk Res, 2010, 34(3): 393～398
37. Lusis E A, Watson M A, Chicoine M R, et al. Integrative genomic analysis identifies NDRG2 as a candidate tumor suppressor gene frequentlyinactivated in clinically aggressive meningioma. Cancer Res, 2005, 65(16): 7121～7126
38. Lorentzen A, Vogel L K, Lewinsky R H, et al. Expression of NDRG2 is down-regulated in high-risk adenomas and colorectal carcinoma. BMC Cancer, 2007, 7: 192
39. Choi S C, Yoon S R, Park Y P, et al. Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death. Exp Mol Med, 2007, 39(6): 705～714
40. Hu X L, Liu X P, Lin S X, et al. NDRG2 expression and mutation in human liver and pancreatic cancers. World J Gastroenterol, 2004, 10(23): 3518～3521
41. Felsberg J, Yan P S, Huang T H, et al. DNA methylation and allelic losses on chromosome arm 14q in oligodendroglial tumours. Neuropathol Appl Neurobiol, 2006, 32(5): 517～524
42. Hummerich L, Muller R, Hess J, et al. Identification of novel tumour-associated genes differentially expressed in the process of squamous cell cancer development. Oncogene, 2006, 25(1): 111～121
43. Shi H, Li N, Li S, et al. Expression of NDRG2 in esophageal squamous cell carcinoma. Cancer Sci, 2010,
44. Schilling S H, Hjelmeland A B, Radiloff D R, et al. NDRG4 is required for cell cycle progression and survival in glioblastoma cells. J Biol Chem, 2009, 284(37): 25160～25169
45. Melotte V, Lentjes M H, van den Bosch S M, et al. N-Myc downstream-regulated gene 4 (NDRG4): a candidate tumor suppressor gene and potential biomarker for colorectal cancer. J Natl Cancer Inst, 2009, 101(13): 916～927
46. Zhang J, Li F, Liu X, et al. The repression of human differentiation-related gene NDRG2 expression by Myc via Miz-1-dependent interaction with the NDRG2 core promoter. J Biol Chem, 2006, 281(51): 39159～39168
47. Zhang J, Chen S, Zhang W, et al. Human differentiation-related gene NDRG1 is a Myc downstream-regulated gene that is repressed by Myc on the core promoter region. Gene, 2008, 417(1-2): 5～12
48. Li J, Kretzner L. The growth-inhibitory Ndrg1 gene is a Myc negative target in human neuroblastomas and other cell types with overexpressed N- or c-myc. Mol Cell Biochem, 2003, 250(1-2): 91～105
49. Liu N, Wang L, Li X, et al. N-Myc downstream-regulated gene 2 is involved in p53-mediated apoptosis. Nucleic Acids Res, 2008, 36(16): 5335～5349
50. Pan J, Zhang S, Chen C, et al. Effect of Recombinant Adenovirus-p53 Combined With Radiotherapy on Long-Term Prognosis of Advanced Nasopharyngeal Carcinoma. Journal of Clinical Oncology, 2009, 27(5): 799～804
51. Svensson E, Vidovic K, Olofsson T, et al. The Wilms' tumor gene 1 (WT1) induces expression of the N-myc downstream regulated gene 2 (NDRG2). DNA Cell Biol, 2007, 26(8): 589～597
52. Nichols N R. Ndrg2, a novel gene regulated by adrenal steroids and antidepressants, is highly expressed in astrocytes. Ann N Y Acad Sci, 2003, 1007: 349～356
53. Zhang J, Liu J, Li X, et al. The physical and functional interaction of NDRG2 with MSP58 in cells. Biochem Biophys Res Commun, 2007, 352(1): 6～11
54. Kim Y J, Yoon S Y, Kim J T, et al. NDRG2 expression decreases withtumor stages and regulates TCF/beta-catenin signaling in human colon carcinoma. Carcinogenesis, 2009, 30(4): 598～605
55. Liu N, Wang L, Liu X, et al. Promoter methylation, mutation, and genomic deletion are involved in the decreased NDRG2 expression levels in several cancer cell lines. Biochem Biophys Res Commun, 2007, 358(1): 164～169
56. Tepel M, Roerig P, Wolter M, et al. Frequent promoter hypermethylation and transcriptional downregulation of the NDRG2 gene at 14q11.2 in primary glioblastoma. Int J Cancer, 2008, 123(9): 2080～2086
57. Lin W, Zhang J, Zhang J, et al. RNAi-mediated inhibition of MSP58 decreases tumor growth, migration and invasion in a human glioma cell line. J Cell Mol Med, 2008,
58. Shi H, Chen S, Jin H, et al. Downregulation of MSP58 inhibits growth of human colorectal cancer cells via regulation of the cyclin D1-cyclin-dependent kinase 4-p21 pathway. Cancer Sci, 2009, 100(9): 1585～1590
59. Herzig M, Savarese F, Novatchkova M, et al. Tumor progression induced by the loss of E-cadherin independent of beta-catenin/Tcf-mediated Wnt signaling. Oncogene, 2007, 26(16): 2290～2298
60. Burchfield J G, Lennard A J, Narasimhan S, et al. Akt mediates insulin-stimulated phosphorylation of Ndrg2: evidence for cross-talk with protein kinase C theta. J Biol Chem, 2004, 279(18): 18623～18632
61. Boulkroun S, Fay M, Zennaro M C, et al. Characterization of rat NDRG2 (N-Myc downstream regulated gene 2), a novel early mineralocorticoid-specific induced gene. J Biol Chem, 2002, 277(35): 31506～31515
62. Wielputz M O, Lee I H, Dinudom A, et al. (NDRG2) stimulatesamiloride-sensitive Na+ currents in Xenopus laevis oocytes and fisher rat thyroid cells. J Biol Chem, 2007, 282(38): 28264～28273
63. Mitchelmore C, Buchmann-Moller S, Rask L, et al. NDRG2: a novel Alzheimer's disease associated protein. Neurobiol Dis, 2004, 16(1): 48～58
64. Ezzeddine Z D, Martuza R L, Platika D, et al. Selective killing of glioma cells in culture and in vivo by retrovirus transfer of the herpes simplex virus thymidine kinase gene. New Biol, 1991, 3(6): 608～614
65. Ram Z, Culver K W, Oshiro E M, et al. Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells. Nat Med, 1997, 3(12): 1354～1361
66. Yuan X, Hu J, Belladonna M L, et al. Interleukin-23-expressing bone marrow-derived neural stem-like cells exhibit antitumor activity against intracranial glioma. Cancer Res, 2006, 66(5): 2630～2638
67. Tanaka T, Manome Y, Wen P, et al. Viral vector-mediated transduction of a modified platelet factor 4 cDNA inhibits angiogenesis and tumor growth. Nat Med, 1997, 3(4): 437～442
68. Ke L D, Fueyo J, Chen X, et al. A novel approach to glioma gene therapy: down-regulation of the vascular endothelial growth factor in glioma cells using ribozymes. Int J Oncol, 1998, 12(6): 1391～1396
69. Robinson R. RNAi therapeutics: how likely, how soon?. PLoS Biol, 2004, 2(1): E28
70. Zhang Y, Chen Z D, Du CJ, et al. siRNA targeting survivin inhibits growth and induces apoptosis in human renal clear cell carcinoma 786-O cells. Pathol Res Pract, 2009, 205(12): 823～827
71. Pan J J, Zhang S W, Chen C B, et al. Effect of recombinant adenovirus-p53 combined with radiotherapy on long-term prognosis of advancednasopharyngeal carcinoma. J Clin Oncol, 2009, 27(5): 799～804
72.黄强,浦佩玉,夏之柏, et al.抑癌基因p53增强胶质瘤自杀基因作用的实验研究.中华显微外科杂志, 2009, 32(1)
73. Aghi M, Rabkin S, Martuza R L. Oncolytic herpes simplex virus mutants exhibit enhanced replication in glioma cells evading temozolomide chemotherapy through deoxyribonucleic acid repair. Clin Neurosurg, 2006, 53: 65～76
74.何黎民,董艳,卢亦成.胶质瘤基因治疗的研究进展.第二军医大学学报, 2002, 23(5)
75.杨学军,杨树源.中枢神经系统分类:临床神经外科学诊断分析与治疗要领.杨国瑞.北京:人民军医出版社, 2004161
76.牛峰,邓艳春,秦娜, et al.人NDRG2两亚型基因原核表达载体的构建及表达与纯化.中华神经外科疾病杂志, 2009, 8(2): 135～139
77.尚学义.稳定转染抑癌候选基因ndrg2对脑胶质瘤细胞系抑制作用的研究:[第四军医大学, 2007
78.张璟. Ndrg2相互作用分子筛选及其在NB4白血病细胞分化中表达相关性的研究: [2005
79. Hehir K M, Baguisi A, Pennington S E, et al. A potential antitumor peptide therapeutic derived from antineoplastic urinary protein. Peptides, 2004, 25(4): 543～549
80.张翊. NDRG-2在肿瘤细胞中的初步功能研究: [2004
81. Louis D N, Ohgaki H, Wiestler O D, et al. The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol, 2007, 114(2): 97～109
82.祝源,黄应桥,邹安琪.胶质瘤的分级和预后分析.实用放射学杂志SYFS, 2007, (03)
83. Friedrich M, Villena-Heinsen C, Reitnauer K, et al. Malignancies of the uterine corpus and immunoreactivity score of the DNA "mismatch-repair" enzyme human Mut-S-homologon-2. J Histochem Cytochem, 1999, 47(1): 113～118
84. Mckeever P E. Insights about brain tumors gained through immunohistochemistry and in situ hybridization of nuclear and phenotypic markers. J Histochem Cytochem, 1998, 46(5): 585～594
85. Buckner J C, Brown P D, O'Neill B P, et al. Central nervous system tumors. Mayo Clin Proc, 2007, 82(10): 1271～1286
86. Vecht C J. Effect of age on treatment decisions in low-grade glioma. J Neurol Neurosurg Psychiatry, 1993, 56(12): 1259～1264
87.葛培林,钟云萍, Pei-Lin G E, et al.低级别脑胶质瘤预后分析.肿瘤防治研究, 2005, 32(7)
2. Okuda T, Kondoh H. Identification of new genes ndr2 and ndr3 which are related to Ndr1/RTP/Drg1 but show distinct tissue specificity and response to N-myc. Biochem Biophys Res Commun, 1999, 266(1): 208～215
3. Kokame K, Kato H, Miyata T. Homocysteine-respondent genes in vascular endothelial cells identified by differential display analysis. GRP78/BiP and novel genes. J Biol Chem, 1996, 271(47): 29659～29665
4. van Belzen N, Dinjens W N, Diesveld M P, et al. A novel gene which is up-regulated during colon epithelial cell differentiation and down-regulated in colorectal neoplasms. Lab Invest, 1997, 77(1): 85～92
5. Lin T M, Chang C. Cloning and characterization of TDD5, an androgen target gene that is differentially repressed by testosterone and dihydrotestosterone. Proc Natl Acad Sci U S A, 1997, 94(10): 4988～4993
6. Zhou D, Salnikow K, Costa M. Cap43, a novel gene specifically induced by Ni2+ compounds. Cancer Res, 1998, 58(10): 2182～2189
7.邓艳春,药立波,刘新平, et al.人脑内-含有ACP样结构域新基因的发现.生物化学与生物物理进展, 2001, 28(1): 72～76
8. Nagase T, Ishikawa K, Suyama M, et al. Prediction of the coding sequences of unidentified human genes. XIII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res, 1999, 6(1): 63～70
9. Zhao W, Tang R, Huang Y, et al. Cloning and expression pattern of thehuman NDRG3 gene. Biochim Biophys Acta, 2001, 1519(1-2): 134～138
10. Zhou R H, Kokame K, Tsukamoto Y, et al. Characterization of the human NDRG gene family: a newly identified member, NDRG4, is specifically expressed in brain and heart. Genomics, 2001, 73(1): 86～97
11. Qu X, Jia H, Garrity D M, et al. Ndrg4 is required for normal myocyte proliferation during early cardiac development in zebrafish. Dev Biol, 2008, 317(2): 486～496
12. Krauter-Canham R, Bronner R, Evrard J, et al. A transmitting tissue-and pollen-expressed protein from sunflower with sequence similarity to the human RTP protein. Plant Sci, 1997, 129: 191～202
13. Qu X, Zhai Y, Wei H, et al. Characterization and expression of three novel differentiation-related genes belong to the human NDRG gene family. Mol Cell Biochem, 2002, 229(1-2): 35～44
14. Shaw E, Mccue L A, Lawrence C E, et al. Identification of a novel class in the alpha/beta hydrolase fold superfamily: the N-myc differentiation-related proteins. Proteins, 2002, 47(2): 163～168
15. Okuda T, Kokame K, Miyata T. Differential expression patterns of NDRG family proteins in the central nervous system. J Histochem Cytochem, 2008, 56(2): 175～182
16. Hou W G, Zhao Y, Shen L, et al. Differential expression of N-Myc downstream regulated gene 2 (NDRG2) in the rat testis during postnatal development. Cell Tissue Res, 2009, 337(2): 257～267
17. Hou W, Hu J, Li Y, et al. Altered Expression of NDRG2 in the Testes of Experimental Rat Model of Cryptorchidism. Urology, 2010,
18. Lachat P, Shaw P, Gebhard S, et al. Expression of NDRG1, a differentiation-related gene, in human tissues. Histochem Cell Biol, 2002,118(5): 399～408
19. Wang W, Li Y, Li Y, et al. NDRG3 is an androgen regulated and prostate enriched gene that promotes in vitro and in vivo prostate cancer cell growth. Int J Cancer, 2009, 124(3): 521～530
20. Deng Y, Yao L, Chau L, et al. N-Myc downstream-regulated gene 2 (NDRG2) inhibits glioblastoma cell proliferation. Int J Cancer, 2003, 106(3): 342～347
21.胡晓兰,药立波,张远强, et al. NDRG2在人胚胎组织中的表达分布特点.生理学报, 2006, 58(04): 331～336
22. Shen L, Zhao Z Y, Wang Y Z, et al. Immunohistochemical detection of Ndrg2 in the mouse nervous system. Neuroreport, 2008, 19(9): 927～931
23. Hu X L, Liu X P, Deng Y C, et al. Expression analysis of the NDRG2 gene in mouse embryonic and adult tissues. Cell Tissue Res, 2006, 325(1): 67～76
24. Wang L, Liu N, Yao L, et al. NDRG2 is a new HIF-1 target gene necessary for hypoxia-induced apoptosis in A549 cells. Cell Physiol Biochem, 2008, 21(1-3): 239～250
25. Sugiki T, Murakami M, Taketomi Y, et al. N-myc downregulated gene 1 is a phosphorylated protein in mast cells. Biol Pharm Bull, 2004, 27(5): 624～627
26. Unoki M, Nakamura Y. Growth-suppressive effects of BPOZ and EGR2, two genes involved in the PTEN signaling pathway. Oncogene, 2001, 20(33): 4457～4465
27. Guan R J, Ford H L, Fu Y, et al. Drg-1 as a differentiation-related, putative metastatic suppressor gene in human colon cancer. Cancer Res, 2000, 60(3): 749～755
28. Bandyopadhyay S, Pai S K, Gross S C, et al. The Drg-1 gene suppresses tumor metastasis in prostate cancer. Cancer Res, 2003, 63(8): 1731～1736
29. Bandyopadhyay S, Pai S K, Hirota S, et al. Role of the putative tumor metastasis suppressor gene Drg-1 in breast cancer progression. Oncogene, 2004, 23(33): 5675～5681
30. Bandyopadhyay S, Wang Y, Zhan R, et al. The tumor metastasis suppressor gene Drg-1 down-regulates the expression of activating transcription factor 3 in prostate cancer. Cancer Res, 2006, 66(24): 11983～11990
31. Kovacevic Z, Richardson D R. The metastasis suppressor, Ndrg-1: a new ally in the fight against cancer. Carcinogenesis, 2006, 27(12): 2355～2366
32. Ellen T P, Ke Q, Zhang P, et al. NDRG1, a growth and cancer related gene: regulation of gene expression and function in normal and disease states. Carcinogenesis, 2008, 29(1): 2～8
33. Piquemal D, Joulia D, Balaguer P, et al. Differential expression of the RTP/Drg1/Ndr1 gene product in proliferating and growth arrested cells. Biochim Biophys Acta, 1999, 1450(3): 364～373
34. Angst E, Sibold S, Tiffon C, et al. Cellular differentiation determines the expression of the hypoxia-inducible protein NDRG1 in pancreatic cancer. Br J Cancer, 2006, 95(3): 307～313
35. Gomez-Casero E, Navarro M, Rodriguez-Puebla M L, et al. Regulation of the differentiation-related gene Drg-1 during mouse skin carcinogenesis. Mol Carcinog, 2001, 32(2): 100～109
36. Tschan M P, Shan D, Laedrach J, et al. NDRG1/2 expression is inhibited in primary acute myeloid leukemia. Leuk Res, 2010, 34(3): 393～398
37. Lusis E A, Watson M A, Chicoine M R, et al. Integrative genomic analysis identifies NDRG2 as a candidate tumor suppressor gene frequentlyinactivated in clinically aggressive meningioma. Cancer Res, 2005, 65(16): 7121～7126
38. Lorentzen A, Vogel L K, Lewinsky R H, et al. Expression of NDRG2 is down-regulated in high-risk adenomas and colorectal carcinoma. BMC Cancer, 2007, 7: 192
39. Choi S C, Yoon S R, Park Y P, et al. Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death. Exp Mol Med, 2007, 39(6): 705～714
40. Hu X L, Liu X P, Lin S X, et al. NDRG2 expression and mutation in human liver and pancreatic cancers. World J Gastroenterol, 2004, 10(23): 3518～3521
41. Felsberg J, Yan P S, Huang T H, et al. DNA methylation and allelic losses on chromosome arm 14q in oligodendroglial tumours. Neuropathol Appl Neurobiol, 2006, 32(5): 517～524
42. Hummerich L, Muller R, Hess J, et al. Identification of novel tumour-associated genes differentially expressed in the process of squamous cell cancer development. Oncogene, 2006, 25(1): 111～121
43. Shi H, Li N, Li S, et al. Expression of NDRG2 in esophageal squamous cell carcinoma. Cancer Sci, 2010,
44. Schilling S H, Hjelmeland A B, Radiloff D R, et al. NDRG4 is required for cell cycle progression and survival in glioblastoma cells. J Biol Chem, 2009, 284(37): 25160～25169
45. Melotte V, Lentjes M H, van den Bosch S M, et al. N-Myc downstream-regulated gene 4 (NDRG4): a candidate tumor suppressor gene and potential biomarker for colorectal cancer. J Natl Cancer Inst, 2009, 101(13): 916～927
46. Zhang J, Li F, Liu X, et al. The repression of human differentiation-related gene NDRG2 expression by Myc via Miz-1-dependent interaction with the NDRG2 core promoter. J Biol Chem, 2006, 281(51): 39159～39168
47. Zhang J, Chen S, Zhang W, et al. Human differentiation-related gene NDRG1 is a Myc downstream-regulated gene that is repressed by Myc on the core promoter region. Gene, 2008, 417(1-2): 5～12
48. Li J, Kretzner L. The growth-inhibitory Ndrg1 gene is a Myc negative target in human neuroblastomas and other cell types with overexpressed N- or c-myc. Mol Cell Biochem, 2003, 250(1-2): 91～105
49. Liu N, Wang L, Li X, et al. N-Myc downstream-regulated gene 2 is involved in p53-mediated apoptosis. Nucleic Acids Res, 2008, 36(16): 5335～5349
50. Pan J, Zhang S, Chen C, et al. Effect of Recombinant Adenovirus-p53 Combined With Radiotherapy on Long-Term Prognosis of Advanced Nasopharyngeal Carcinoma. Journal of Clinical Oncology, 2009, 27(5): 799～804
51. Svensson E, Vidovic K, Olofsson T, et al. The Wilms' tumor gene 1 (WT1) induces expression of the N-myc downstream regulated gene 2 (NDRG2). DNA Cell Biol, 2007, 26(8): 589～597
52. Nichols N R. Ndrg2, a novel gene regulated by adrenal steroids and antidepressants, is highly expressed in astrocytes. Ann N Y Acad Sci, 2003, 1007: 349～356
53. Zhang J, Liu J, Li X, et al. The physical and functional interaction of NDRG2 with MSP58 in cells. Biochem Biophys Res Commun, 2007, 352(1): 6～11
54. Kim Y J, Yoon S Y, Kim J T, et al. NDRG2 expression decreases withtumor stages and regulates TCF/beta-catenin signaling in human colon carcinoma. Carcinogenesis, 2009, 30(4): 598～605
55. Liu N, Wang L, Liu X, et al. Promoter methylation, mutation, and genomic deletion are involved in the decreased NDRG2 expression levels in several cancer cell lines. Biochem Biophys Res Commun, 2007, 358(1): 164～169
56. Tepel M, Roerig P, Wolter M, et al. Frequent promoter hypermethylation and transcriptional downregulation of the NDRG2 gene at 14q11.2 in primary glioblastoma. Int J Cancer, 2008, 123(9): 2080～2086
57. Lin W, Zhang J, Zhang J, et al. RNAi-mediated inhibition of MSP58 decreases tumor growth, migration and invasion in a human glioma cell line. J Cell Mol Med, 2008,
58. Shi H, Chen S, Jin H, et al. Downregulation of MSP58 inhibits growth of human colorectal cancer cells via regulation of the cyclin D1-cyclin-dependent kinase 4-p21 pathway. Cancer Sci, 2009, 100(9): 1585～1590
59. Herzig M, Savarese F, Novatchkova M, et al. Tumor progression induced by the loss of E-cadherin independent of beta-catenin/Tcf-mediated Wnt signaling. Oncogene, 2007, 26(16): 2290～2298
60. Burchfield J G, Lennard A J, Narasimhan S, et al. Akt mediates insulin-stimulated phosphorylation of Ndrg2: evidence for cross-talk with protein kinase C theta. J Biol Chem, 2004, 279(18): 18623～18632
61. Boulkroun S, Fay M, Zennaro M C, et al. Characterization of rat NDRG2 (N-Myc downstream regulated gene 2), a novel early mineralocorticoid-specific induced gene. J Biol Chem, 2002, 277(35): 31506～31515
62. Wielputz M O, Lee I H, Dinudom A, et al. (NDRG2) stimulatesamiloride-sensitive Na+ currents in Xenopus laevis oocytes and fisher rat thyroid cells. J Biol Chem, 2007, 282(38): 28264～28273
63. Mitchelmore C, Buchmann-Moller S, Rask L, et al. NDRG2: a novel Alzheimer's disease associated protein. Neurobiol Dis, 2004, 16(1): 48～58
64. Ezzeddine Z D, Martuza R L, Platika D, et al. Selective killing of glioma cells in culture and in vivo by retrovirus transfer of the herpes simplex virus thymidine kinase gene. New Biol, 1991, 3(6): 608～614
65. Ram Z, Culver K W, Oshiro E M, et al. Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells. Nat Med, 1997, 3(12): 1354～1361
66. Yuan X, Hu J, Belladonna M L, et al. Interleukin-23-expressing bone marrow-derived neural stem-like cells exhibit antitumor activity against intracranial glioma. Cancer Res, 2006, 66(5): 2630～2638
67. Tanaka T, Manome Y, Wen P, et al. Viral vector-mediated transduction of a modified platelet factor 4 cDNA inhibits angiogenesis and tumor growth. Nat Med, 1997, 3(4): 437～442
68. Ke L D, Fueyo J, Chen X, et al. A novel approach to glioma gene therapy: down-regulation of the vascular endothelial growth factor in glioma cells using ribozymes. Int J Oncol, 1998, 12(6): 1391～1396
69. Robinson R. RNAi therapeutics: how likely, how soon?. PLoS Biol, 2004, 2(1): E28
70. Zhang Y, Chen Z D, Du CJ, et al. siRNA targeting survivin inhibits growth and induces apoptosis in human renal clear cell carcinoma 786-O cells. Pathol Res Pract, 2009, 205(12): 823～827
71. Pan J J, Zhang S W, Chen C B, et al. Effect of recombinant adenovirus-p53 combined with radiotherapy on long-term prognosis of advancednasopharyngeal carcinoma. J Clin Oncol, 2009, 27(5): 799～804
72.黄强,浦佩玉,夏之柏, et al.抑癌基因p53增强胶质瘤自杀基因作用的实验研究.中华显微外科杂志, 2009, 32(1)
73. Aghi M, Rabkin S, Martuza R L. Oncolytic herpes simplex virus mutants exhibit enhanced replication in glioma cells evading temozolomide chemotherapy through deoxyribonucleic acid repair. Clin Neurosurg, 2006, 53: 65～76
74.何黎民,董艳,卢亦成.胶质瘤基因治疗的研究进展.第二军医大学学报, 2002, 23(5)
75.杨学军,杨树源.中枢神经系统分类:临床神经外科学诊断分析与治疗要领.杨国瑞.北京:人民军医出版社, 2004161
76.牛峰,邓艳春,秦娜, et al.人NDRG2两亚型基因原核表达载体的构建及表达与纯化.中华神经外科疾病杂志, 2009, 8(2): 135～139
77.尚学义.稳定转染抑癌候选基因ndrg2对脑胶质瘤细胞系抑制作用的研究:[第四军医大学, 2007
78.张璟. Ndrg2相互作用分子筛选及其在NB4白血病细胞分化中表达相关性的研究: [2005
79. Hehir K M, Baguisi A, Pennington S E, et al. A potential antitumor peptide therapeutic derived from antineoplastic urinary protein. Peptides, 2004, 25(4): 543～549
80.张翊. NDRG-2在肿瘤细胞中的初步功能研究: [2004
81. Louis D N, Ohgaki H, Wiestler O D, et al. The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol, 2007, 114(2): 97～109
82.祝源,黄应桥,邹安琪.胶质瘤的分级和预后分析.实用放射学杂志SYFS, 2007, (03)
83. Friedrich M, Villena-Heinsen C, Reitnauer K, et al. Malignancies of the uterine corpus and immunoreactivity score of the DNA "mismatch-repair" enzyme human Mut-S-homologon-2. J Histochem Cytochem, 1999, 47(1): 113～118
84. Mckeever P E. Insights about brain tumors gained through immunohistochemistry and in situ hybridization of nuclear and phenotypic markers. J Histochem Cytochem, 1998, 46(5): 585～594
85. Buckner J C, Brown P D, O'Neill B P, et al. Central nervous system tumors. Mayo Clin Proc, 2007, 82(10): 1271～1286
86. Vecht C J. Effect of age on treatment decisions in low-grade glioma. J Neurol Neurosurg Psychiatry, 1993, 56(12): 1259～1264
87.葛培林,钟云萍, Pei-Lin G E, et al.低级别脑胶质瘤预后分析.肿瘤防治研究, 2005, 32(7)
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |