OBJECTIVETo study the thymidine phosphorylase (TP) expression in different types of cancer and its correlation with tumor microvessel density (MVD).

METHODSThe expression of TP and MVD was detected by immunohistochemistry method. In a series of 251 cancer patients there were 48 patients with gastric cancer, 53 with colorectal cancer, 47 with breast cancer, 56 with cervical cancer, 47 with lung cancer. Normal gastric (n = 25), colorectal (n = 25), cervical (n = 17) and lung (n = 25) tissues around the cancer were also examined.

RESULTSThe TP expression rate was 64.6% in gastric cancer, 67.9% in colorectal cancer, 80.9% in breast cancer, 82.1% in cervical cancer, and 63.8% in lung cancer, which was significantly higher than that in normal tissues (P = 0.0000). TP expression was positively correlated with MVD in gastric, colorectal, breast, and cervical cancers. The correlation was not statistically significant in lung cancer.

CONCLUSIONThis study indicates that TP overexpression in cancer may be associated with tumor angiogenesis.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; blood supply ; enzymology ; Colorectal Neoplasms ; blood supply ; enzymology ; Female ; Humans ; Male ; Middle Aged ; Neovascularization, Pathologic ; pathology ; Stomach Neoplasms ; blood supply ; enzymology ; Thymidine Phosphorylase ; metabolism ; Uterine Cervical Neoplasms ; blood supply ; enzymology

OBJECTIVETo investigate the transcription level and protein expression of HIF-1alpha and VEGF in SW480 cell line and colorectal adenocarcinoma, and to determine whether HIF-1alpha plays a role in angiogenesis through its regulation of VEGF.

METHODSHIF-1alpha mRNA expression was analyzed by in situ hybridization. HIF-1alpha and VEGF protein expressions were determined by immunochemical streptavidin/peroxidase (SP) in SW480 cells and colorectal carcinoma tissue samples and Western blot, using proteins extracted from SW480 cells. Tumor tissue microvessel density (MVD) was determined by CD34 immunostaining of colorectal carcinomas.

RESULTSThe levels of HIF-1alpha mRNA changed significantly in response to different oxygen concentrations and an addition of genistein in SW480 cells. Immunocytochemistry revealed that the levels of HIF-1alpha, VEGF protein expression in SW480 cells were significantly higher under hypoxia than those in nomoxia (P < 0.01, P < 0.05 respectively). However, addition of genistein, an inhibitor of HIF-1alpha, suppressed such responses to hypoxia. Western blot analysis showed that SW480 cells exposed to hypoxia expressed a high level of HIF-1alpha protein, compared to a weak expression in nomoxia. The addition of genistein in hypoxia suppressed the over-expression of HIF-1alpha. The positive rates of HIF-1alpha mRNA by in situ hybridization in colorectal adenomas and adenocarcinomas were 38.9% (7/18) and 67.7% (42/62), respectively. The percentage of HIF-1alpha mRNA positive cells varied significantly from colorectal adenomas to adenocarcinomas at different Duke stages (P < 0.05), and HIF-1alpha mRNA was higher in adenocarcinomas than in adenomas (P < 0.01). The positive rates of HIF-1alpha and VEGF protein expression in adenocarcinomas were 43.5% (27/62) and 37.1% (23/62), respectively. The expression of VEGF elevated as the Duke tumor staging increased. The conformation rate of HIF-1alpha and VEGF was 74.2% (46/62). MVD was significantly higher in HIF-1alpha and/or VEGF positive tumors than those without (P < 0.01 and P < 0.05 respectively). Among the four groups, i.e. HIF-1alpha+/VEGF+, HIF-1alpha+/VEGF-, HIF-1alpha+/VEGF- and HIF-1alpha-/VEGF-, the difference of MVD was highly significant (P < 0.01). HIF-1alpha expression was correlated significantly with VEGF expression and microvessel density.

CONCLUSIONSThese findings suggest hypoxia induces the expression of HIF-1alpha and VEGF in colorectal adenocarcinoma. HIF-1alpha may play an important role in angiogenesis and tumor progression by regulating the expression of VEGF in human colorectal carcinoma.

Adenocarcinoma ; blood supply ; metabolism ; pathology ; Colorectal Neoplasms ; blood supply ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Microcirculation ; pathology ; Neovascularization, Pathologic ; etiology ; RNA, Messenger ; biosynthesis ; genetics ; Transcription Factors ; biosynthesis ; genetics ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9), which degrade extracellular matrix, are believed to play a crucial role in tumor invasion and metastasis. Angiogenesis is also perceived as an important step in tumor growth and metastasis. To investigate the expression of MMPs and the correlation between the expression of MMPs and angiogenesis in colorectal adenocarcinoma, we studied 72 cases of colorectal adenocarcinoma in Inha University Hospital from 1996 to 1997. We evaluated the expression of MMPs by immunohistochemistry and angiogenesis by counting the microvessels. The expression of MMP-2 was increased according to the Astler-Coller stage (p< 0.05). Angiogenesis in the metastatic group was higher than that of the localized one (p<0.05). The expression of MMP-2 positively correlated with angiogenesis (p<0.05), and marked expression of MMP-9 positively correlated with angiogenesis (p<0.05). The present results suggest that the expression of MMP-2 provides clues for tumor progression and angiogenesis provides significant information to predict whether metastasis is present in colorectal adenocarcinoma.
Adenocarcinoma/pathology ; Adenocarcinoma/metabolism* ; Adenocarcinoma/blood supply ; Adolescence ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34/biosynthesis ; Collagenases/biosynthesis* ; Colorectal Neoplasms/pathology ; Colorectal Neoplasms/metabolism* ; Colorectal Neoplasms/blood supply ; Gelatinase A ; Gelatinase B ; Gelatinases/biosynthesis* ; Human ; Metalloendopeptidases/biosynthesis* ; Middle Age ; Neovascularization, Pathologic*/pathology

OBJECTIVETo investigate the expression of angiopoietin (Ang)-1 and Ang-2 in colorectal tumors and its relations to microvessel density (MVD) in tumor tissue.

METHODSAng-1, Ang-2 and factor VIII-related antigen were stained immunohistochemically in 91 cases of primary colorectal adenocarcinoma, 20 cases of colorectal adenoma and 24 cases of normal colorectal mucosal tissue, and MVD was also assayed in above tissue specimens.

RESULT(1) A significantly higher Ang-1 (7.07+/-2.00) was observed in normal tissue compared with 1.75 +/-1.98 in the adenoma and 1.40 +/- 1.22 in the adenocarcinoma (P<0.01). (2) Ang-2 protein positive rate in adenocarcinoma was significantly higher than that in normal tissue and adenoma (P<0.01). The expression of Ang-2 in adenocarcinoma was closely associated with poor differentiation and vessel invasion. (3) There were significant correlations between Ang-1 and Ang-2 (r=-0.338, P<0.01), Ang-1 and MVD (r=-0.388, P<0.01), Ang-2 and MVD (r=0.594, P<0.01) in the 135 cases.

CONCLUSIONThe overexpression of Ang-2 may play an important role in angiogenesis of colorectal adenocarcinoma. It can be regarded as an index for malignancy and prognosis in colorectal adenocarcinoma.

Adenocarcinoma ; blood supply ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Angiopoietin-1 ; biosynthesis ; genetics ; Angiopoietin-2 ; biosynthesis ; genetics ; Biomarkers, Tumor ; Capillaries ; pathology ; Colorectal Neoplasms ; blood supply ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; von Willebrand Factor ; biosynthesis ; genetics

OBJECTIVETo investigate the association of the expressions of angiopoietin-2 (Ang-2), vascular endothelial growth factor (VEGF) in colorectal cancer tissues with Dukes' clinicopathological features.

METHODSAng-2 and Tie-2 mRNA expressions were detected in colorectal cancer tissues, adjacent tissues, and normal tissues by real time-PCR. Quantikine immunoassays were used to measure the protein expressions of Ang-2 and VEGF in the tissues and serum samples.

RESULTSAng-2 and Tie-2 levels were significantly higher in the serum of the patients than in the normal tissues (P<0.05), and their expressions were strongly correlated (r=0.879, P=0.000). Tumor tissue Ang-2 and VEGF levels were significantly higher than their levels in the adjacent and normal tissues (P<0.05). In colorectal cancer patients, the peripheral blood level of Ang-2 was significantly higher than that in healthy control subjects, and comparable with that in mesenteric blood (P>0.05). In Dukes' stage C and D patients, serum Ang-2 and VEGF levels were significantly higher than those in patients in Dukes' stage A and B (P<0.05).

CONCLUSIONAng-2 and VEGF over expressions may play an important role in the occurrence and progression of colorectal cancer.

Adult ; Aged ; Angiopoietin-2 ; blood ; metabolism ; Case-Control Studies ; Colorectal Neoplasms ; blood supply ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factor A ; blood ; metabolism

Contrast-enhanced computed tomography colonography (CE-CTC) is a useful guide for the laparoscopic surgeon to avoid incorrectly removing the colonic segment and the failure to diagnose of synchronous colonic and extra-colonic lesions. Lymph node dissection and vessel ligation under a laparoscopic approach can be time-consuming and can damage vessels and organs. Moreover, mesenteric vessels have extreme variations in terms of their courses and numbers. We describe the benefit of using an abdominal vascular map created by CE-CTC in laparoscopic colorectal surgery candidates. We describe patients with different diseases (colorectal cancer, diverticular disease, and inflammatory bowel disease) who underwent CE-CTC just prior to laparoscopic surgery.
Adult ; Aged ; Colectomy/*methods ; Colon/blood supply/pathology/radiography ; Colonography, Computed Tomographic/*methods ; Colorectal Neoplasms/pathology/*radiography/*surgery ; Contrast Media ; Female ; Humans ; Laparoscopy/*methods ; Lymph Node Excision/methods ; Male ; Middle Aged ; Neoplasm Staging/methods

BACKGROUND/AIMS: Lymph node (LN) metastasis occurs in approximately 10% of patients with submucosally invasive colorectal carcinoma. This study was performed to determine the role of matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs) production and microvessel formation on the LN metastasis in submucosally invasive colorectal carcinoma. METHODS: A total of forty-one subjects with surgically resected submucosally invasive colorectal carcinoma were included in this study. Immunohistochemical staining of MMP-2, MMP-9, TIMP-1, TIMP-2, and urokinase-type plasminogen activator were performed. Angiogenesis was evaluated by counting the number of microvessels in each pathologic specimen as identified by CD34 immunohistochemical staining. RESULTS: The depth of submucosal invasion was not significantly correlated with the expression of MMP-2, MMP-9, TIMP-1, TIMP-2, or urokinase-type plasminogen activator, but the microvessel count was significantly correlated with the absolute depth of invasion (r=0.312, p<0.05). Upregulation of TIMP-2 was positively correlated with adjacent lymphatic invasion (p<0.05) and increased TIMP-2 expression was correlated with LN metastasis in submucosally invasive colorectal carcinoma (p=0.088). CONCLUSIONS: These results suggest that the expression of TIMP-2 and the microvessel count may be useful parameters for considering additional surgery after endoscopic treatment of submucosally invasive colorectal carcinoma.
Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms/blood supply/*metabolism/pathology ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinases/*metabolism ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic/*pathology ; Tissue Inhibitor of Metalloproteinases/*metabolism

OBJECTIVE: To investigate the correlation between quantitative dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) parameters and microvascular density (MVD) in a human-colon-cancer xenograft mouse model using 3 Tesla MRI. MATERIALS AND METHODS: A human-colon-cancer xenograft model was produced by subcutaneously inoculating 1 x 106 DLD-1 human-colon-cancer cells into the right hind limbs of 10 mice. The tumors were allowed to grow for two weeks and then assessed using MRI. DCE-MRI was performed by tail vein injection of 0.3 mmol/kg of gadolinium. A region of interest (ROI) was drawn at the midpoints along the z-axes of the tumors, and a Tofts model analysis was performed. The quantitative parameters (Ktrans, Kep and Ve) from the whole transverse ROI and the hotspot ROI of the tumor were calculated. Immunohistochemical microvessel staining was performed and analyzed according to Weidner's criteria at the corresponding MRI sections. Additional Hematoxylin and Eosin staining was performed to evaluate tumor necrosis. The Mann-Whitney test and Spearman's rho correlation analysis were performed to prove the existence of a correlation between the quantitative parameters, necrosis, and MVD. RESULTS: Whole transverse ROI of the tumor showed no significant relationship between the MVD values and quantitative DCE-MRI parameters. In the hotspot ROI, there was a difference in MVD between low and high group of Ktrans and Kep that had marginally statistical significance (ps = 0.06 and 0.07, respectively). Also, Ktrans and Kep were found to have an inverse relationship with MVD (r = -0.61, p = 0.06 in Ktrans; r = -0.60, p = 0.07 in Kep). CONCLUSION: Quantitative analysis of T1-weighted DCE-MRI using hotspot ROI may provide a better histologic match than whole transverse section ROI. Within the hotspots, Ktrans and Kep tend to have a reverse correlation with MVD in this colon cancer mouse model.
Animals ; Capillary Permeability ; Colorectal Neoplasms/*blood supply/pathology ; *Contrast Media ; Female ; Gadolinium/diagnostic use ; Humans ; Image Processing, Computer-Assisted ; Immunohistochemistry ; *Magnetic Resonance Imaging ; Mice ; Mice, Nude ; Microvessels/*pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic/diagnosis

OBJECTIVETo explore the correlation between metabolic tumour volume (MTV) and microvessel density (MVD) and blood-borne metastasis in colorectal carcinoma.

METHODSThirty-six patients with CRC conformed by pathology underwent PET-CT examination before operation. SUVmax and MTV were obtained by PET VCRA software. The blood vessels were identified with CD34 immunohistochemical staining, and the MVD was recorded. The correlation between SUVmax and MTV with histological differentiation, T stage, MVD and blood-borne metastasis was analyzed.

RESULTSThe SUVmax, MTV and MVD in patients with blood-borne metastasis were 5.15 ± 5.41, (22.99 ± 18.63) cm³ and 14.17 ± 3.63, and were 10.65 ± 3.79, (16.95 ± 11.82) cm³ and 11.27 ± 3.69, respectively, in patients with non-blood-borne metastasis. The differences of SUVmax, MTV and MVD between blood-borne metastasis and non-blood-borne metastasis patients were statistically significant (all P > 0.05). Pearson correlation analysis found that there was no linear correlation between SUVmax and MVD, and the SUVmax was not statistically significant between high and low MVD groups (t = 0.919, P = 0.364). But there was a linear correlation between MTV and MVD (r = 0.621, P = 0.000), and the MTV was statistically significant between high and low MVD groups (t = 3.567, P = 0.001). The receiver-operating characteristic curves showed that MTV could be used to predict blood-borne metastasis of CRC, and the best cutoff value for MTV was 14.975 cm³, and the sensitivity, specificity, negative predictive value and positive predictive value were 85.7%, 54.5%, 72.3% and 64.2%, respectively. There were no significant relationships between SUVmax, MTV, MVD, blood-borne metastasis and histological differentiation (P > 0.05). With the increased T stage, the MTV, MVD and the probability of blood-borne metastasis were also increased (all P < 0.05).

CONCLUSIONSThere are correlations between MTV and MVD and blood-borne metastasis in CRC. The risk of blood-borne metastasis in patients with MTV > 14.975 cm³ is higher, and needs to take more effective intervention.

Colorectal Neoplasms ; blood supply ; diagnostic imaging ; pathology ; Fluorodeoxyglucose F18 ; Humans ; Microvessels ; pathology ; Multimodal Imaging ; Neoplasm Staging ; Neoplastic Cells, Circulating ; Positron-Emission Tomography ; ROC Curve ; Radiopharmaceuticals ; Sensitivity and Specificity ; Tomography, X-Ray Computed

BACKGROUNDRNA interference (RNAi) technology is emerging as a very potent tool to generate a cellular knockdown of a desired gene. The aim of this study was to explore whether RNAi targeting vascular endothelial growth factor-C (VEGF-C) could inhibit colorectal tumor lymphangiogenesis and tumor growth.

METHODSWe used vector-based RNAi to inhibit VEGF-C expression in colon cancer in vitro and in vivo. VEGF-C expression was quantified by real-time polymerase chain reaction and Westen blotting. To establish LoVo cell tumor xenografts in mice, we subcutaneously inoculated 1.0 x 10(6) LoVo cells in nude mice; after injection, tumors were allowed to grow for 4 weeks until the volume reached (75.8+/-55.8) mm(3). The mice were then randomly divided into two groups: (1) the VEGF-C-siRNA group (n=10) received direct injection of "therapeutic" plasmid 50 microg of LoVo-VEGF-C-siRNA into the tumor mass; (2) the control group (n=10) were injected with LoVo-control in 20 microl of sterile 0.9% NaCl solution into the tumor mass. Tumor growth, microlymphatics and microvessels were compared for mice administered either systemic VEGF-C-siRNA or control over 4 weeks.

RESULTSThe mRNA and protein expression of VEGF-C in the colon tumor cell line, LoVo, stably transfected with a VEGF-C-siRNA vector, were significantly downregulated 45.3% and 35.3% respectively. In vivo, four weeks after injection, the tumor volume were significantly smaller in VEGF-C-siRNA group than in LoVo-control group ((314.8+/-54.8) mm(3) vs (553.9+/-90.1) mm(3)); the incidences of lymph node metastasis (30%) in VEGF-C-siRNA were significantly inhibited compared with LoVo-control group (70%); in VEGF-C-siRNA group, the number of microlymphatics per microscopic field was (5.3+/-0.7) and the number of microvessels per microscopic field was (24.2+/-6.5) decreased compared with LoVo-control group (12.5+/-6.9) and (42.1+/-7.4) (all P<0.001).

CONCLUSIONInhibition of VEGF-C expression using siRNA-mediated gene silencing vectors reduced lymphangiogenesis and lymph node metastasis and enhanced survival.

Animals ; Cell Line, Tumor ; Colorectal Neoplasms ; blood supply ; pathology ; therapy ; Genetic Vectors ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Mice ; Neovascularization, Pathologic ; prevention & control ; RNA Interference ; RNA, Small Interfering ; genetics ; Vascular Endothelial Growth Factor C ; antagonists & inhibitors ; genetics