Identification of Tumor Angiogenesis-Related Genes by Subtractive Hybridization

Wang, Jing Liang; Liu, Yeheng H.; Lee, Maggie C.; Nguyen, Tri Khoa; Lee, Chris; Kim, Ann; Nguyen, Mai

doi:10.1006/mvre.2000.2242

Cited by 18 publications

(15 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some of the identi®ed genes have already been shown to be regulated in endothelial cells by FGF2, like integrin a 3 (Klein et al, 1993) andvimentin (Carey andZehner, 1995), or by VEGF (thymosin b10, Vasile et al, 2001). Also, the study of gene expression pro®les during angiogenesis in vitro and in vivo has identi®ed several cytoskeletal components, as well as extracellular matrix components and their receptors, as upregulated in activated endothelial cells (Glienke et al, 2000;Kahn et al, 2000;St Croix et al, 2000;Wang et al, 2000). Relevant to this point is the observation that various angiogenesis inhibitors may exert their angiostatic activity by interacting with cytoskeleton (e.g.…”

Section: Arrays (Clontech) a Typical Results Is Reported In (A) And (mentioning

confidence: 99%

“…In the last decade several targets for anti-angiogenic therapy have been discovered leading to the development of angiostatic compounds; some of them, including inhibitors of a v b 3 integrin and of the vascular endothelial growth factor (VEGF) signaling pathway, are currently under test in clinical trials (for a review see Liekens et al, 2001a). In this contest, gene expression pro®les have been described for endothelial cells activated in vitro (Glienke et al, 2000;Kahn et al, 2000;Roland et al, 2000;Wang et al, 2000) or in vivo during tumor angiogenesis (St Croix et al, 2000). The results have allowed the identi®cation of several known and unknown gene transcripts upregulated in endothelial cells under these conditions.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Gene expression profile in fibroblast growth factor 2-transformed endothelial cells

et al. 2002

View full text Add to dashboard Cite

Fibroblast growth factor-2 (FGF2) exerts paracrine and autocrine functions on endothelial cells. FGF2-overexpressing murine aortic endothelial cells (FGF2-T-MAE cells) induce opportunistic hemangioendothelioma-like tumors when inoculated in immunode®cient mice. To evaluate the impact of FGF2-mediated activation on gene expression pro®le in transformed endothelial cells, we performed subtractive suppression hybridization analysis between FGF2-T-MAE cells and parental MAE cells. The two cell populations were compared for di erential gene expression also by gene macroarray hybridization with 32 P-labeled cDNAs. The two approaches allowed the identi®cation of 27 transcripts whose expression was upregulated by FGF2 in endothelial cells. With the exception of one unknown gene, the di erentially expressed transcripts encoded for proteins involved in the modulation of cell cycle, di erentiation, and cell adhesion. Among them, the stress-inducible genes A170, GADD45 and GADD153 are upregulated by FGF2 transfection or recombinant growth factor treatment. Their expression was also induced in vascular tumors originated by parental or FGF2-transfected MAE cells in nude mice. This study extends the number of genes involved in tumor angiogenesis and/or endothelial cell transformation, a ®nding with possible implications for the discovery of novel targets for angiostatic therapy.

show abstract

Section: Arrays (Clontech) a Typical Results Is Reported In (A) And (mentioning

confidence: 99%

mentioning

confidence: 99%

Gene expression profile in fibroblast growth factor 2-transformed endothelial cells

et al. 2002

View full text Add to dashboard Cite

show abstract

“…This approach has been successfully used for tissue-specific cDNA and genomic DNA subtraction (25,26) to isolate cancer-related and differentiation stage-specific genes (27)(28)(29)(30) and to clone drug-induced genes or pathogens from infected tissues (31)(32)(33). As compared with other low-yield subtractive hybridization techniques, the experimental approach used in this study is applicable to small amounts of rare samples, such as RNA extracted from islets derived from patients with recent onset of diabetes.…”

Section: Discussionmentioning

confidence: 99%

A Reg Family Protein Is Overexpressed in Islets From a Patient With New-Onset Type 1 Diabetes and Acts as T-Cell Autoantigen in NOD Mice

Gürr

Yavari

et al. 2002

Diabetes

View full text Add to dashboard Cite

Genes overexpressed in pancreatic islets of patients with new-onset type 1 diabetes are potential candidates for novel disease-related autoantigens. RT-PCR-based subtractive hybridization was used on islets from a patient who died at the onset of type 1 diabetes, and it identified a type 1 diabetes-related cDNA encoding hepatocarcinoma-intestine-pancreas/pancreatic-associated protein (HIP/PAP). This protein belongs to the family of Reg proteins implicated in islet regeneration; its gene contains a putative interleukin-6 (IL-6) response element. Islets from healthy cadaveric human donors released HIP/PAP protein into the culture medium, and this release was enhanced by the addition of IL-6. The expression pattern of mouse homologues of HIP/PAP was determined in pancreata of prediabetic and diabetic NOD mice. Both groups showed positive immunostaining for HIP/PAP in islets and ductal epithelium. To test whether HIP/PAP is a target of isletdirected autoimmunity, we measured splenic T-cell responses against HIP/PAP in NOD mice. Spontaneous proliferation was detected after 4 weeks. Lymphocytes from islet infiltrates and pancreatic lymph nodes from 7-to 10-week-old NOD mice were used to establish an HIP/PAP-specific I-A g7 -restricted T-cell line, termed WY1, that also responded to mouse islets. WY1 cells homed to islets of NOD-SCID mice and adoptively transferred disease when coinjected with purified CD8 ؉ cells from diabetic NOD mice. Our conclusion was that differential cloning of Reg from islets of a type 1 diabetic patient and the response of Reg to the cytokine IL-6 suggests that HIP/PAP becomes overexpressed in human diabetic islets because of the local inflammatory response. HIP/PAP acts as a T-cell autoantigen in NOD mice. Therefore, autoimmunity to HIP/PAP might create a vicious cycle, accelerating the immune process leading to diabetes. Diabetes 51:339 -346, 2002

show abstract

“…Toward this goal, we used a subtraction hybridization method called suppression subtractive hybridization (12). In human umbilical vein endothelial cell populations exposed to conditioned media from human cancer cells (13) for 4 hours, we have isolated multiple clones (14,15). One of these differentially expressed genes is endothelial-derived gene 1 (EG-1; ref.…”

Section: Introductionmentioning

confidence: 99%

The Novel Gene EG-1 Stimulates Cellular Proliferation

Lu¹,

Zhang²,

Maul³

et al. 2005

Cancer Research

View full text Add to dashboard Cite

We recently discovered a novel gene and named it endothelialderived gene 1 (EG-1). Previously, we have shown that the expression of EG-1 is significantly elevated in the epithelial cells of breast cancer, colorectal cancer, and prostate cancer. Here, we report that EG-1 can stimulate cellular proliferation. Transfection experiments which overexpressed the full-length EG-1 gene in human embryonic kidney HEK-293 cells or human breast cancer cell lines resulted in significantly increased in vitro proliferation, in comparison with transfection with empty vectors. On the other hand, small interfering RNA cotransfection resulted in inhibition of proliferation. S.c. xenograft assays were carried out in a severe combined immunodeficient mouse model. We found that injection of high EG-1 expressing HEK-293 clones resulted in significantly larger tumors, in comparison with clones carrying the empty vectors. To further clarify the function of this gene, we investigated its interaction with Src and members of the mitogen-activated protein kinase (MAPK) family. Immunoprecipitation with anti-Src antibody, followed by immunoblotting with anti-EG-1 antibody, showed an association between these two molecules. Overexpression of EG-1 was correlated with activation of the following kinases: extracellular signal-regulated kinases 1 and 2, c-jun-NH 2 -kinase, and p38. These observations collectively support the hypothesis that the novel gene EG-1 is a positive stimulator of cellular proliferation, and may possibly be involved in signaling pathways involving Src and MAPK activation. (Cancer Res 2005; 65(14): 6159-66)

show abstract

Identification of Tumor Angiogenesis-Related Genes by Subtractive Hybridization

Cited by 18 publications

References 7 publications

Gene expression profile in fibroblast growth factor 2-transformed endothelial cells

Gene expression profile in fibroblast growth factor 2-transformed endothelial cells

A Reg Family Protein Is Overexpressed in Islets From a Patient With New-Onset Type 1 Diabetes and Acts as T-Cell Autoantigen in NOD Mice

The Novel Gene EG-1 Stimulates Cellular Proliferation

Contact Info

Product

Resources

About