OBJECTIVE: To explore the expression of vascular endothelial growth factor (VEGF) C and D in gastric cancer and its relationship with tumor angiogenesis and lymph node metastasis. METHODS: Immunohistochemistry(SABC) and real-time PCR were used to detect the expression of VEGF-C, VEGF-D protein and mRNA in 32 gastric cancer tissues and 32 normal gastric tissues. RESULTS: The positive expression rate of VEGF-C and VEGF-D in gastric cancer tissue was significantly higher than that of normal gastric tissues (P<0.01), the expression of VEGF-C and VEGF-D in the gastric cancer group and the lymph node metastasis group were significantly different (P<0.05). The expression of VEGF-C and VEGF-D in gastric carcinoma was positively correlated with lymph node metastasis (P<0.01), the expression of VEGF-C and VEGF-D in well-differentiated carcinoma, moderately differentiated carcinoma and poorly differentiated carcinoma was statistically different (P<0.05). VEGF-C and VEGF-D expressions in gastric cancer cells were not related to the patient's age, sex, and lymph node distant metastasis (P>0.05). CONCLUSION The non-intake high expression of VEGF-C and VEGF-D in gastric cancer cells is closely related to lymph node metastasis. They serve as the important reference indicator to assess the prognosis in gastric cancer patients.
Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; Vascular Endothelial Growth Factor D ; genetics ; metabolism

BACKGROUNDHyaluronidase (Hyase) is an enzyme which hydrolyses hyaluronan (HA), a large nonsulfated glycosaminoglycan. Several genes have been identified to code for hyaluronidases in humans. Its role has only recently been underlined in the invasion of prostate cancer, colonic cancer, and breast cancer. Moreover, the findings were in agreement with some experimental results which showed that HA-derived oligosaccharides had angiogenesis-promoting activity. All these findings prompted us to investigate factors that had been characterized as putative invasive factors in different human breast cancer-derived cell lines.

METHODSWe selected two series of human breast cancer-derived cell lines whose expression of estrogen receptors (ER) was previously published. Hyaluronidase secretion in culture medium and expression of matrix metallo-proteinase (MMP)-9, cathepsin-D (cath-D) and vascular endothelial growth factor (VEGF) by cells were determined. We also investigated cell invasiveness in the Matrigel invasion assay, and studied the capability of cancer cells to promote in vitro formation of tubules by endothelial cells.

RESULTSER(-) cells secreted significantly more hyaluronidase (P < 0.001) and expressed significantly more VEGF (P < 0.01), MMP-9 (P < 0.05) and cath-D (P < 0.0001) than ER(+) cells. Invasion through Matrigel by ER(-) Hyase(+) cells was significantly higher than that by ER(+) Hyase(-) cells (P < 0.05). In both cases, invasion was decreased by heparin (P < 0.05). When ECV-304 endothelial cells were co-cultivated in millicell chambers with cancer cells, ECV-304 cells were induced to form tubules. Tubule formation was demonstrated to be more prominent with ER(-) Hyase(+) cells than with ER(+) Hyase(-) cells (P < 0.05).

CONCLUSIONInvasive features of ER(-) breast cancer cells can be characterized in vitro by an invasive Matrigel assay, as the induction of tubule formation by ECV-304 endothelial cells, higher secretion of hyaluronidase, and higher expression of proteinases MMP-9, cath-D, and the angiogenesis promoting factor VEGF.

Breast Neoplasms ; metabolism ; Cathepsin D ; metabolism ; Cell Line, Tumor ; Humans ; Hyaluronoglucosaminidase ; metabolism ; Immunohistochemistry ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; genetics ; Receptors, Estrogen ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

Growth factor-stimulated phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine (PC), generating phosphatidic acid (PA) which may act as a second messenger during cell proliferation and survival. Therefore, PLD is believed to play an important role in tumorigenesis. In this study, a potential mechanism for PLD-mediated tumorigenesis was explored. Ectopic expression of PLD1 or PLD2 in human glioma U87 cells increased the expression of hypoxia-inducible factor-1alpha (HIF-1alpha) protein. PLD-induced HIF-1 activation led to the secretion of vascular endothelial growth factor (VEGF), a HIF-1 target gene involved in tumorigenesis. PLD induction of HIF-1alpha was significantly attenuated by 1-butanol which blocks PA production by PLD, and PA per se was able to elevate HIF-1alpha protein level. Inhibition of mTOR, a PA-responsive kinase, reduced the levels of HIF-1alpha and VEGF in PLD-overexpressed cells. Epidermal growth factor activated PLD and increased the levels of HIF-1alpha and VEGF in U87 cells. A specific PLD inhibitor abolished expression of HIF-1alpha and secretion of VEGF. PLD may utilize HIF-1-VEGF pathway for PLD-mediated tumor cell proliferation and survival.
Cell Line, Tumor ; Epidermal Growth Factor/metabolism ; Gene Expression Regulation, Neoplastic ; Glioma/genetics/*metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics/metabolism ; Phosphatidic Acids/*metabolism ; Phospholipase D/genetics/*metabolism ; *Signal Transduction ; Transfection ; Vascular Endothelial Growth Factor A/*metabolism

OBJECTIVETo study mRNA expression of VEGF-(A, C, D) genes in breast carcinoma and its effect on prognosis.

METHODSThe mRNA expression of VEGF-(A, C, D) genes in 61 cases of primary breast carcinoma and 29 cases of benign mammary lesions was detected by TaqMan real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) technology. The correlation between mRNA expression and various clinicopathologic parameters and survival data was analyzed.

RESULTSThe level of mRNA expression of VEGF-(A, C) genes was significantly higher in primary breast carcinomas (2.79 +/- 1.31 and 3.33 +/- 0.88 respectively) than in benign mammary lesions (1.59 +/- 1.35 and 2.76 +/- 0.55 respectively, P = 0.000 and 0.002 respectively). The percentage of VEGF-D mRNA expression was 73.77% in breast carcinomas, as compared to 51.72% in benign mammary lesions (P = 0.038). However, there was no statistically significant difference in the mRNA expression levels of VEGF-D gene between primary breast carcinomas and benign mammary lesions (P = 0.683). On the other hand, the ratio of VEGF-D mRNA expression to VEGF-C mRNA expression was lower in lymph node-positive than in lymph node-negative cases. This also correlated with the occurrence of nodal metastasis by uni- and multivariate analysis (P(uni) = 0.046 and P(mult) = 0.062). High mRNA expression of VEGF-(A, C) genes was associated with poor disease-free survival (P = 0.030 and 0.044 respectively).

CONCLUSIONSThe expression of VEGF-(A, C, D) genes may play a role in the disease progression of breast carcinoma. The ratio of VEGF-D mRNA expression to VEGF-C mRNA expression correlates with the occurrence of lymph node metastasis in breast carcinoma. The mRNA expression of VEGF-(A, C) genes may serve as a useful prognostic indicator in breast cancer.

Adult ; Aged ; Breast Neoplasms ; genetics ; pathology ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Kaplan-Meier Estimate ; Middle Aged ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; genetics ; Vascular Endothelial Growth Factor C ; genetics ; Vascular Endothelial Growth Factor D ; genetics

Cardiac lymphatic system in the remodeling after acute myocardial infarction (AMI) has been overlooked. We wanted to investigate the role of bone marrow-derived endothelial progenitor cells (EPCs) and their contribution to lymphatic distribution in myocardial remodeling after AMI. Mouse (C57bl/6J) MI models were created by ligation of the left anterior descending coronary artery and were treated with phosphate buffered saline (PBS) or EPCs. Real-time RT-PCR with 2- to 4-week myocardial tissue samples revealed that lymphangiogenetic factors such as vascular endothelial growth factor (VEGF)-C (8.5 fold, P < 0.05), VEGF-D (6.1 fold, P < 0.05), Lyve-1 (15 fold, P < 0.05), and Prox-1 (11 fold, P < 0.05) were expressed at significantly higher levels in the PBS group than the EPC group. The PBS group also showed a significantly higher density of lymphatic vessels in the peri-infarction area. Echocardiography showed that from 2 weeks after the treatment, left ventricle (LV) dimensions at both systole and diastole were significantly smaller in the EPC group than in the PBS group (P < 0.01) and LV fractional shortening was higher in the EPC group accordingly (P < 0.01). Lymphangiogenic markers increased in a mouse MI model. EPC transplantation decreased lymphangiogenesis and adverse ventricular remodeling after AMI. These novel findings suggest that new lymphatic vessels may be formed in severely damaged myocardium, and may be involved in adverse myocardial remodeling after AMI.
Animals ; Cell Transplantation ; Endothelial Cells/*cytology ; Homeodomain Proteins/genetics/metabolism ; Immunohistochemistry ; Lymphangiogenesis/genetics/*physiology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Myocardial Infarction/metabolism/physiopathology/*therapy ; Real-Time Polymerase Chain Reaction ; *Stem Cell Transplantation ; Tumor Suppressor Proteins/genetics/metabolism ; Vascular Endothelial Growth Factor A/genetics/metabolism ; Vascular Endothelial Growth Factor D/genetics/metabolism

OBJECTIVETo investigate the correlation of vascular endothelial growth factor D (VEGF-D) expression, microlymphatic density (MLD) and microvessel density (MVD) levels with the development and metastasis of rectal cancer.

METHODSEighty specimens from resected middle-lower rectal cancer diagnosed by pathology were examined by immunohistochemistry for VEGF-D,MLD and MVD. Simultaneously, 40 biopsy specimens from rectal polyps and 80 specimens from normal rectal tissue were examined as controls. Correlation between the expression of above three factors and the tumor size, gross morphology, histological type, metastasis, differentiation grade, infiltration depth, Dukes stage, lymph node metastasis and long-distance metastasis before operation were investigated with Spearman method.

RESULTS(1) Positive expression rate of VEGF-D was 55 % (44/80) in rectal cancer, and zero in rectal polyps and normal rectal tissues. The expression of VEGF-D in rectal cancer was significantly higher than that in rectal polyps and normal rectal tissues(P<0.05). MLD was significantly higher in rectal cancer (2.80+/-1.31) than that in rectal polyps (0.50+/-0.72) and normal rectal tissues(0.25+/-0.44)(P<0.05).Meanwhile MVD was significantly higher in rectal cancer (80.10+/-23.18) than that in rectal polyps (27.00+/-11.01) and normal rectal tissues (10.45+/-5.34) (P<0.05). (2) VEGF-D, MLD and MVD were positively correlated with lymph node metastasis and long-distance metastasis before operation (P<0.05). (3) VEGF-D was positively correlated with MLD (P<0.05) and MLD was positively correlated with MVD as well(P<0.05).

CONCLUSIONSLymphangiogenesis exists in rectal cancer tissues. VEGF-D and MLD can be used as good predictors of lymphangiogenesis and they are the important factors affecting biological behavior of rectal cancer. Lymphangiogenesis and angiogenesis may have a cooperative function in the development of rectal cancer.

Female ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Microcirculation ; Microvessels ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; pathology ; Rectal Neoplasms ; blood supply ; pathology ; Vascular Endothelial Growth Factor D ; genetics ; metabolism