OBJECTIVETo investigate intratumoral microvessel density (MVD) and expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (BFGF) in hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE) and to evaluate their significance.

METHODSMVD and expression of VEGF and BFGF in cancerous tissues were examined in forty specimens resected from patients with HCC using immunohistochemical methods. Among these patients, 20 patients received 1 to 7 treatments of TACE prior to II-phase surgical resection (TACE group), the other 20 patients were treated by operation without receiving any other treatment preoperatively (surgical group). There was no significant difference in clinical features between the two groups. MVD was assessed by counting immunostained endothelial cells within a certain area, and staining intensity of VEGF was assessed quantitatively with computer-assisted image analyzer. The expression of BFGF was determined by cell-positive or cell-negative.

RESULTSThe average MVD was 130.51 75.5 in TACE group and 152.35 58.80 in surgical group. There was no significant difference between the two groups (t=-1.021, P=0.341). Staining intensity of VEGF was 645.60 543.27 in TACE group, higher than in surgical group (158.28 188.48, t=281, P<0.001). BFGF-positive rate was 35% in TACE group and 40% in surgical group. There was no significant difference (x(2)=0.107, P=0.744).

CONCLUSIONSThe results indicate that survived cancerous tissue has rich vascularity and the expression of VEGF of the cancerous cells can be enhanced by TACE which may play an important role in reestablishment of blood supply to tumor after TACE.

Carcinoma, Hepatocellular ; metabolism ; pathology ; physiopathology ; therapy ; Catheterization ; Embolization, Therapeutic ; Endothelial Growth Factors ; biosynthesis ; Fibroblast Growth Factor 2 ; biosynthesis ; Humans ; Liver Neoplasms ; metabolism ; pathology ; physiopathology ; therapy ; Lymphokines ; biosynthesis ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

To investigate the effect of vascular endothelial growth factor (VEGF) on beta1 integrin (VLA-4 and VLA-5) activation ability in K562 cells transfected with antisense VEGF121 cDNA, K562 cells were transfected with antisense (As), sense (S) and vector (V, pcDNA(3)). Flow cytometry was used to evaluate the expression rate of VLA-4 (CD49d/CD29) and VLA-5 (CD49e/CD29) and beta1 integrin activation ability in the transfected K562 cells. The results showed that the expression rates of VLA-4 and VLA-5 were more than 92% in the transfected K562 cells and there was no difference among the K562/V, K562/S and K562/As cells. However, beta1 integrin activated 9EG7 expression rate in K562/As cells was higher than that in K562/V cells [(75.6 +/- 10.5)% vs (41.2 +/- 2.1)%, P < 0.01)] after activation with beta1 integrin activator 8A2. It is concluded that function of beta1 integrin to be activated is restored in K562 cells transfected with antisense VEGF121 cDNA.
DNA ; genetics ; DNA, Antisense ; genetics ; Endothelial Growth Factors ; genetics ; metabolism ; Flow Cytometry ; Humans ; Integrin alpha4beta1 ; biosynthesis ; Integrin alpha5beta1 ; biosynthesis ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Lymphokines ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the the expression and hypoxic regulation of vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9.

METHODSVEGF mRNA and MMP-9 mRNA were examined by reverse transcription-polymerase chain reaction (RT-PCR) in 43 esophageal carcinoma specimens including 18 para-tumorous esophageal tissues. The expression of VEGF protein and mean microvessel density (MVD) in 56 specimens were examined by immunohistochemical stain. The effect of hypoxia on VEGF and MMP-9 expression in esophageal cancer cell lines was quantitatively determined by enzyme linked immunosorbent assay (ELISA).

RESULTSThe VEGF expression in the tumorous tissue, being significantly correlated with MVD in the tumor, was remarkably higher than that in the para-tumorous tissue. VEGF and MVD expression in the tumor was significantly associated with stage and metastasis of esophageal carcinoma. The MMP-9 expression in the tumorous tissue, being uncorrelated with vessel count and clinicopathologic features in esophageal carcinoma, was significantly higher than that in the para-tumorous tissue. Hypoxia significantly increased the VEGF expression in esophageal cancer cell lines but did not affect the MMP-9 expression.

CONCLUSIONSThe expression of VEGF plays an important role in the angiogenesis and metastasis of esophageal cancer, which is regulated by hypoxia. VEGF may serve as a predictor of progression in esophageal carcinoma and a potential target for antiangiogenic therapy of esophageal carcinoma.

Endothelial Growth Factors ; biosynthesis ; genetics ; Esophageal Neoplasms ; metabolism ; Female ; Gene Expression Regulation ; Humans ; Hypoxia ; Lymphokines ; biosynthesis ; genetics ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Middle Aged ; Oxygen ; metabolism ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

To investigate the relationship of the expression of vascular endothelial factor (VEGF) and matrix metalloproteinase-2 (MMP-2) with the recurrence and metastasis of hepatocellular carcinoma (HCC), the expression of VEGF and MMP-2 in HCC tissue(n = 50) and in normal liver tissue(n = 30) were examined by immunochemistry. The results showed that the positive rates of VEGF and MMP-2 in HCC tissue were 86% and 60% respectively, and in normal liver tissue were 53.3% and 30% respectively. The positive rates of VEGF and MMP-2 in HCC were significantly higher than those in normal liver tissue. The positive rates of VEGF and MMP-2 in HCC with intra- or extra-hepatic metastasis were higher than those of HCC without metastasis. VEGF and MMP-2 play important roles in the invasion and metastasis of HCC.
Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; secondary ; Endothelial Growth Factors ; biosynthesis ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Liver Neoplasms ; metabolism ; pathology ; Lymphokines ; biosynthesis ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

To investigate the relationship of the expression of vascular endothelial growth factor (VEGF) and C-myc with the occurrence, development and metastasis of gallbladder carcinoma, the expression levels of VEGF and C-myc in gallbladder carcinoma tissue (n = 30) and in normal gallbladder tissue (n = 20) were examined by immunochemistry. Results show that the positive rates of VEGF and C-myc in gallbladder carcinoma tissue were 80% and 63.3% respectively, and 45% and 25% respectively in normal gallbladder tissue. The positive rates of VEGF and C-myc were significantly higher in gallbladder carcinoma than in normal gallbladder tissue. The expression of VEGF and C-myc in gallbladder carcinoma related to the metastasis of gallbladder carcinoma. VEGF and C-myc play important roles in the occurrence, development and metastasis of gallbladder carcinoma.
Adult ; Aged ; Endothelial Growth Factors ; biosynthesis ; physiology ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; physiology ; Lymphokines ; biosynthesis ; physiology ; Male ; Middle Aged ; Neoplasm Metastasis ; Proto-Oncogene Proteins c-myc ; biosynthesis ; physiology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Vascular endothelial growth factor (VEGF) is known to play a key role in tumor angiogenesis. The tumor-suppressor gene p53 has been thought to regulate VEGF. We investigated the effect of VEGF on esophageal carcinoma and the correlation between VEGF and p53. Tissue samples were taken from 81 patients with esophageal carcinoma after surgery. VEGF and p53 expressions were examined by immunohistochemical staining. Microvessels in the tumor stained for CD34 antigen were also counted. VEGF and p53 expressions were observed in 51.3% (41/80) and 51.9% (41/79), respectively. The microvessel density was 70.9+/-6.7 (mean+/-SE) in VEGF-positive group and 68.7+/-5.1 in VEGF-negative group. However, no correlation was noted between VEGF and p53 expression. Whereas the tumor size, nodal status, depth of invasions, and tumor stage were associated with poor overall survival, VEGF expression or p53 expression was not. These results indicate that VEGF and p53 are highly expressed in esophageal carcinomas. Since the VEGF expression is not correlated with the p53 expression, microvessel density or clinicopathological findings, further studies with other angiogenic molecules are needed to determine the role in esophageal carcinomas.
Adult ; Aged ; Capillaries ; Carcinoma, Squamous Cell/classification/*metabolism/pathology ; Endothelial Growth Factors/*biosynthesis ; Esophageal Neoplasms/classification/*metabolism/pathology ; Female ; Humans ; Lymphatic Metastasis ; Lymphokines/*biosynthesis ; Male ; Middle Aged ; Neoplasm Staging ; *Neovascularization, Pathologic ; Prognosis ; Retrospective Studies ; Tumor Suppressor Protein p53/*biosynthesis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Mesenchymal stem cells (MSCs) can inhibit T cell proliferation; however, the underlying mechanisms are not clear. In this study, we investigated the mechanisms of the immunoregulatory activity of MSCs on T cells. Irradiated MSCs co-cultured with either naive or pre-activated T cells in a mixed lymphocyte reaction (MLR) significantly suppressed T cell proliferation in a dose-dependent manner, irrespective of allogeneic disparity between responders and MSCs. Transwell assays revealed that the suppressive effect was primarily mediated by soluble factors that induced apoptosis. Splenocytes stimulated with alloantigen in the presence of the MSC culture supernatant (CS) produced a significant amount of IL-10, which was attributed to an increase in the number of IL-10 secreting cells, confirmed by an ELISPOT assay. The blockade of IL-10 and IL-10 receptor interaction by anti-IL-10 or anti-IL-10-receptor antibodies abrogated the suppressive capacity of MSC CS, indicating that IL-10 plays a major role in the suppression of T cell proliferation. The addition of 1-methyl-DL-tryptophan (1-MT), an indoleamine 2,3-dioxygenase (IDO) inhibitor, also restored the proliferative capacity of T cells. In conclusion, we demonstrated that soluble mediators from culture supernatant of MSCs could suppress the proliferation of both naive and pre-activated T cells in which IL-10 and IDO play important roles.
Animals ; Cell Proliferation ; Cells, Cultured ; Indoleamine-Pyrrole 2,3,-Dioxygenase/antagonists & inhibitors/metabolism ; Interleukin-10/*biosynthesis ; *Lymphocyte Activation ; Lymphokines/pharmacology ; Mesenchymal Stem Cells/cytology/*metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Interleukin-10/metabolism ; T-Lymphocytes/cytology/*immunology/metabolism ; Tryptophan/analogs & derivatives/pharmacology

Mesenchymal stem cells (MSCs) can inhibit T cell proliferation; however, the underlying mechanisms are not clear. In this study, we investigated the mechanisms of the immunoregulatory activity of MSCs on T cells. Irradiated MSCs co-cultured with either naive or pre-activated T cells in a mixed lymphocyte reaction (MLR) significantly suppressed T cell proliferation in a dose-dependent manner, irrespective of allogeneic disparity between responders and MSCs. Transwell assays revealed that the suppressive effect was primarily mediated by soluble factors that induced apoptosis. Splenocytes stimulated with alloantigen in the presence of the MSC culture supernatant (CS) produced a significant amount of IL-10, which was attributed to an increase in the number of IL-10 secreting cells, confirmed by an ELISPOT assay. The blockade of IL-10 and IL-10 receptor interaction by anti-IL-10 or anti-IL-10-receptor antibodies abrogated the suppressive capacity of MSC CS, indicating that IL-10 plays a major role in the suppression of T cell proliferation. The addition of 1-methyl-DL-tryptophan (1-MT), an indoleamine 2,3-dioxygenase (IDO) inhibitor, also restored the proliferative capacity of T cells. In conclusion, we demonstrated that soluble mediators from culture supernatant of MSCs could suppress the proliferation of both naive and pre-activated T cells in which IL-10 and IDO play important roles.
Animals ; Cell Proliferation ; Cells, Cultured ; Indoleamine-Pyrrole 2,3,-Dioxygenase/antagonists & inhibitors/metabolism ; Interleukin-10/*biosynthesis ; *Lymphocyte Activation ; Lymphokines/pharmacology ; Mesenchymal Stem Cells/cytology/*metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Interleukin-10/metabolism ; T-Lymphocytes/cytology/*immunology/metabolism ; Tryptophan/analogs & derivatives/pharmacology

Leptin, the product of ob gene, is an endocrine hormone that regulates adipose tissue mass. Recently, leptin has been found to generate a growth signal involving a tyrosine kinase-dependent intracellular pathway and promote angiogenic processes via activation of leptin receptor (Ob-R) in endothelial cells. However, it is not clear how leptin functions to promote multi-step processes involved in the neovascularization at the atherosclerotic plaque. We have examined the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) and Ob-R in human atherosclerotic lesions, leptin-mediated angiogenesis in vivo and in vitro. Immunohistochemical analysis of human atherosclerotic aorta revealed an increased expression of Ob-R in the intima of neorevascularized regions and of both MMPs and TIMPs predominantly in the endothelial lining of intimal neovessels and macrophages/foam cells. In the rat corneal angiogenesis assay, leptin elicited a comparable sensitivity of angiogenic activity to those of vascular endothelial growth factor (VEGF). The immunohistological analysis of the leptin-treated rat cornea showed definitive rises in Ob-R, MMPs and TIMPs expression as well as those of VEGF receptor (VEGFR-1). Leptin (10-40 ng/ml) induced proliferation of the human umbilical vein endothelial cells (HUVECs) and elevation of MMP-2, MMP-9, TIMP-1, and TIMP-2 expression in a dose-dependent manner. Leptin also induced increases of MMP-2, MMP-9, TIMP-1, and Up-regulated the human coronary artery smooth muscle cells (HCASMCs). These findings suggest that leptin, a hormone with pluralistic properties including a mitogenic activity on vascular endothelial cells, plays a role in matrix remodeling by regulating the expression of MMPs and TIMPs. Taken together, our findings further provide evidences for leptin's role as an angiogenesis inducer in the normal organ (rat cornea) and in aberrant vasculature under duress like atherosclerosis.
Animal ; Arteriosclerosis/metabolism ; Blotting, Western ; Cell Division ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Growth Factors/metabolism ; Endothelium, Vascular/*cytology/*enzymology ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2/metabolism ; Immunohistochemistry ; Leptin/*chemistry/metabolism/*physiology ; Lymphokines/metabolism ; Matrix Metalloproteinases/*biosynthesis ; *Neovascularization, Pathologic ; Rats ; Receptor Protein-Tyrosine Kinases/metabolism ; Receptors, Growth Factor/metabolism ; Recombinant Proteins/metabolism ; Support, Non-U.S. Gov't ; Tissue Inhibitor of Metalloproteinases/metabolism ; Umbilical Veins/metabolism ; Up-Regulation