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Identification of novel targets for antiangiogenic therapy by comparing the gene expressions of tumor and normal endothelial cells.

Otsubo T, Hida Y, Ohga N, Sato H, Kai T, Matsuki Y, Takasu H, Akiyama K, Maishi N, Kawamoto T, Shinohara N, Nonomura K, Hida K - Cancer Sci. (2014)

Bottom Line: Targeting tumor angiogenesis is an established strategy for cancer therapy.We identified 131 genes that were differentially upregulated in mTEC.The expression of DEF6 and TMEM176B was upregulated in tumor vessels of human renal cell carcinoma specimens, suggesting that they are potential targets for antiangiogenic intervention for renal cell carcinoma.

View Article: PubMed Central - PubMed

Affiliation: Drug Discovery II, DSP Cancer Institute, Dainippon Sumitomo Pharma Co., Ltd, Osaka, Japan.

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Related in: MedlinePlus

Characterization of murine tumor endothelial cells (mTEC) and murine normal endothelial cells (mNEC). (a) The expression of Cd31, Cd105, Vegfr1, Vegfr2, Cd11b, Cd45 and human HBEGF in mTEC (Melanoma-EC, Renal Ca-EC and Oral Ca-EC) and mNEC (Skin-EC) was analyzed using RT-PCR. CD31-negative non-EC fractions and human tumor cells (melanoma, renal carcinoma and oral carcinoma) were also analyzed. (b) Isolated and cultured mEC formed tubes on matrigel-coated plates. Bar, 100 μm. EC, endothelial cells; Renal Ca-EC, Renal carcinoma-EC; Oral Ca-EC, Oral carcinoma-EC; mEC, murine endothelial cells.
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fig01: Characterization of murine tumor endothelial cells (mTEC) and murine normal endothelial cells (mNEC). (a) The expression of Cd31, Cd105, Vegfr1, Vegfr2, Cd11b, Cd45 and human HBEGF in mTEC (Melanoma-EC, Renal Ca-EC and Oral Ca-EC) and mNEC (Skin-EC) was analyzed using RT-PCR. CD31-negative non-EC fractions and human tumor cells (melanoma, renal carcinoma and oral carcinoma) were also analyzed. (b) Isolated and cultured mEC formed tubes on matrigel-coated plates. Bar, 100 μm. EC, endothelial cells; Renal Ca-EC, Renal carcinoma-EC; Oral Ca-EC, Oral carcinoma-EC; mEC, murine endothelial cells.

Mentions: We first isolated and cultured three different types of mTEC (Melanoma-EC, Renal carcinoma-EC [Renal Ca-EC] and Oral carcinoma-EC [Oral Ca-EC]) from human tumor xenografts and mNEC (Skin-EC) from dermal tissues of nude mice as a normal control. These murine endothelial cells (mEC) were positive for the EC markers Cd31, Cd105, Vegfr1 and Vegfr2 and negative for the monocyte marker Cd11b and hematopoietic marker Cd45 using RT-PCR (Fig.1a). Human HBEGF, which is expressed in human tumor cells, was not detected in any of the mTEC (Fig.1a). These results excluded the possibility that these mEC were contaminated with non-EC such as monocytes, hematopoietic cells and human tumor cells. Furthermore, tube formation was observed when mEC were cultured on matrigel-coated plates (Fig.1b), indicating that these mEC maintained EC properties after isolation and culture. Thus, our isolation technique yielded highly pure and functional populations of mEC suitable for subsequent analyses.


Identification of novel targets for antiangiogenic therapy by comparing the gene expressions of tumor and normal endothelial cells.

Otsubo T, Hida Y, Ohga N, Sato H, Kai T, Matsuki Y, Takasu H, Akiyama K, Maishi N, Kawamoto T, Shinohara N, Nonomura K, Hida K - Cancer Sci. (2014)

Characterization of murine tumor endothelial cells (mTEC) and murine normal endothelial cells (mNEC). (a) The expression of Cd31, Cd105, Vegfr1, Vegfr2, Cd11b, Cd45 and human HBEGF in mTEC (Melanoma-EC, Renal Ca-EC and Oral Ca-EC) and mNEC (Skin-EC) was analyzed using RT-PCR. CD31-negative non-EC fractions and human tumor cells (melanoma, renal carcinoma and oral carcinoma) were also analyzed. (b) Isolated and cultured mEC formed tubes on matrigel-coated plates. Bar, 100 μm. EC, endothelial cells; Renal Ca-EC, Renal carcinoma-EC; Oral Ca-EC, Oral carcinoma-EC; mEC, murine endothelial cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317838&req=5

fig01: Characterization of murine tumor endothelial cells (mTEC) and murine normal endothelial cells (mNEC). (a) The expression of Cd31, Cd105, Vegfr1, Vegfr2, Cd11b, Cd45 and human HBEGF in mTEC (Melanoma-EC, Renal Ca-EC and Oral Ca-EC) and mNEC (Skin-EC) was analyzed using RT-PCR. CD31-negative non-EC fractions and human tumor cells (melanoma, renal carcinoma and oral carcinoma) were also analyzed. (b) Isolated and cultured mEC formed tubes on matrigel-coated plates. Bar, 100 μm. EC, endothelial cells; Renal Ca-EC, Renal carcinoma-EC; Oral Ca-EC, Oral carcinoma-EC; mEC, murine endothelial cells.
Mentions: We first isolated and cultured three different types of mTEC (Melanoma-EC, Renal carcinoma-EC [Renal Ca-EC] and Oral carcinoma-EC [Oral Ca-EC]) from human tumor xenografts and mNEC (Skin-EC) from dermal tissues of nude mice as a normal control. These murine endothelial cells (mEC) were positive for the EC markers Cd31, Cd105, Vegfr1 and Vegfr2 and negative for the monocyte marker Cd11b and hematopoietic marker Cd45 using RT-PCR (Fig.1a). Human HBEGF, which is expressed in human tumor cells, was not detected in any of the mTEC (Fig.1a). These results excluded the possibility that these mEC were contaminated with non-EC such as monocytes, hematopoietic cells and human tumor cells. Furthermore, tube formation was observed when mEC were cultured on matrigel-coated plates (Fig.1b), indicating that these mEC maintained EC properties after isolation and culture. Thus, our isolation technique yielded highly pure and functional populations of mEC suitable for subsequent analyses.

Bottom Line: Targeting tumor angiogenesis is an established strategy for cancer therapy.We identified 131 genes that were differentially upregulated in mTEC.The expression of DEF6 and TMEM176B was upregulated in tumor vessels of human renal cell carcinoma specimens, suggesting that they are potential targets for antiangiogenic intervention for renal cell carcinoma.

View Article: PubMed Central - PubMed

Affiliation: Drug Discovery II, DSP Cancer Institute, Dainippon Sumitomo Pharma Co., Ltd, Osaka, Japan.

Show MeSH
Related in: MedlinePlus