ObjectiveTo investigate the expression of keratinocyte growth factor (KGF) and cyclooxygen-ase-2 (COX-2) protein and microvessel density (MVD), and to explore their function and mechanism in the multistep process of gastric cancer. MethodsThe expressions of KGF and COX-2 protein in 64 samples of gastric cancer and 30 cases of normal gastric mucosa tissues were detected by immunohistochemistry. The MVD was detected by staining the endothelial cells in microvessles using anti-CD34 antibody. ResultsThe positive rate of KGF and COX-2 protein expression in gastric cancer were 65.6% (42/64) and 79.7% (51/64), respectively, which was significantly higher than that in normal gastric mucosa tissues 〔(23.3%, 7/30), P=0.046; (13.3%, 4/30), P=0.008〕. The MVD of gastric cancer was 31.8±8.0, which was significantly higher than that of normal gastric mucosa tissues (14.3±6.1), P=0.000. The MVD in gastric cancer with coexpressive KGF and COX-2 protein was 35.9±5.7, which was significant higher than that with non-coexpressive KGF and COX-2 protein (25.7±7.0), P=0.000. Both the expression of KGF and COX-2 protein were related to the invasion of serosa, lymph node metastasis and TNM staging (Plt;0.05, Plt;0.01). The MVD of gastric cancer tissues was related to lymph node metastasis and TNM staging (Plt;0.05), but unrelated to patient’s age, gender, and differentiation of tumor (Pgt;0.05). The co-expression of KGF and COX-2 protein was frequently found in patients with deeper invasion of serosa, lymph node metastasis, and higher TNM staging (Plt;0.05), but which was not associated withpatient’sage, gender, and differentiation of tumor (Pgt;0.05). The expression of KGF protein was positively correlated to the expression of COX-2 protein (r=0.610, P=0.000). There was positive correlation between MVD and the expression of KGF (r=0.675, P=0.000) and COX-2 protein (r=0.657, P=0.000) in gastric cancer, respectively. ConclusionKGF and COX-2 highly expressed by gastric cancer, which may be involved in the invasion and metastasis of gastric cancer by synergisticly promoting the angiogenesis.
ObjectiveTo explore the relation between vascular endothelial growth factor （VEGF） and the formation of tumor thrombosis in the main trunks of portal vein (PVTT). MethodsTumor specimens were collected from 36 patients (16 patients with PVTT, the other patients without PVTT and metastasis) undergoing resection of hepatocellular carcinoma (HCC) and portal thrombectemy, PVTT specimens of 16 patients named group A1, the same patients’ with HCC named group A2, tumor specimens of the other patients named group B. In situ hybridization and immunohistochemistry were used to investigate VEGF mRNA, protein and microvessel density (MVD) on surgical specimens. The intensity was evaluated using a computer image analyzercell analysis system.ResultsVEGF mRNA expression was detected in the tumor’ cell of the specimens. The expression rates of VEGF mRNA in the group B, A2, A1 were 30%, 100%, 100% respectively, and the expression rates of VEGF mRNA in group A2 and A1 were higher than that in group B （P＜0.01）. The intensity of VEGF mRNA in group A2 （0.078 5±0.019 6） were lower than in group A1 （0.194 4±0.059 0） （P＜0.01）. VEGF protein expression was often detected in the tumor cell, vascular endothelial cell and fibroblast cells. Invasion was detected in small vein in group A2, more tumor cell colony detected in group A1. The expression rates of VEGF protein in group B, A2, A1 were same as VEGF mRNA; the intensity of VEGF protein in A1 （0.165 6± 0.034 5） was higher than in group A2 （0.108 1±0.024 3） （P＜0.01）. MVD in group B, A2, A1 was 31.9±14.4, 63.3±15.1, 116±27.6/view of 200 microscopefield, MVD in group A1 was higher than group A2 （P＜0.01）, higher in group A2 than in group B. There was a statistically significant correlation between the intensity of VEGF expression and MVD in group B,A2 and A1. ConclusionVEGF could play an important role in the invasion, metastasis of HCC and the formation of PVTT. Angiogenesis in tumor is correlated well with the progression of HCC.
【Abstract】Objective To investigate the correlation between the expression of fibronectin (FN) in extracellular matrix (ECM) and angiogenesis of gastric carcinoma.Methods The expressions of FN and vascular endothelial growth factor (VEGF) in 20 specimens of normal gastric tissue (normal group) and 80 specimens of gastric carcinoma tissue(gastric carcinoma group) were detected by EnVisonTM immunohistochemical technique. Tumor microvessel densit y (MVD) was evaluated by using antiCD34 antibody as an endothelial marker by the same technique as well. Results The immune complex of FN stained in brown were distributed around glands and in connective tissue of gastric specimens. In normal group, the staining of FN formed intact linear structure at basement membrane and presented regular striae form in connective tissue. In gastric carcinoma group, the integrity of linear structure of FN staining at basement membrane were destroyed to different extent and the staining of FN in connective tissue were changed deeper and distributed irregularly. The expression of VEGF and the value of MVD in the gastric carcinoma group was higher than those in normal group’s(P＜0.01, P＜0.01, respectively).This study indicated, that in gastric carcinoma group, the degree of FN expression in connective tissue had statistically positive correlations with the degree of VEGF expression and MVD value(P＜0.01, P＜0.05, respectively). Whereas the destruction extent of linear structure of FN staining at basement membrane showed no correlation with VEGF expression and MVD value(Pgt;0.05, Pgt;0.05, respectively).Conclusion The higher expression of FN in connective tissue of gastric carcinoma may well play a critical role in its process of angiogenesis and vasculogenesis. There may be an cooperative interactions between FN and VEGF in the process of angiogenesis and vasculogenesis of gastric carcinoma.
Objective To observe the expression levels of nuclear factor kappa B （NF-κB）， vascular endothelial growth factor （VEGF）， and CD31 in portal vein and surrounding tissues of rats during the formation process of cavernoustransformation of portal vein （CTPV）， and try to search the relationship between NF-κB， VEGF， and the angiogenesisof portal areas， as well as the significance and the role of NF-κB and VEGF in the formation process of CTPV. Methods One hundred and ten Sprague-Dawley （SD） rats were randomly （random number method） divided into sham operation group and model group. The partial constriction operations on portal vein were performed in model rats with a blunt 21Gcaliber to establish CTPV animal models （model group）， while the exploratory operations on portal vein， not constriction，were performed in rats of sham operation group. All specimens （portal vein and surrounding tissues） were fixed in formalinand made into paraffin blocks. Each specimen was tested by immunohistochemistry for the expressions of NF-κB， VEGF， and CD31， then optical density （OD） of NF-κB expression and the mean integral optical density （IOD） of VEGF expressionwere measured by using Image Pro Plus 6.0 software， and microvessel density （MVD） was calculated under microscope. Results Nucleoplasm ratio of OD value of NF-κB， mean IOD value of VEGF， and MVD value in 1， 2， 3， 4， and 6 weeks after operation didn’t significantly differed from that of before operation in sham operation group （P＞0.05）， but higher at all time points after operation in model group （P＜0.01）. Compared with sham operation group， nucleoplasm ratio of OD value of NF-κB， mean IOD value of VEGF， and MVD value were significantly higher in 1， 2， 3， 4， and 6 weeks after operation in model group （P＜0.01）. NF-κB and VEGF， NF-κB and MVD， VEGF and MVD were positively correlated with each other （r=0.654 6，P＜0.01；r=0.620 7， P＜0.01；r=0.636 9， P＜0.01） in model group. Conclusion NF-κB and VEGF may relate to the formation of CTPV， and may involve in the angiogenesis.
Objective To study the expression of inducible nitric oxide synthase (iNOS)，endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) in human gastric cancer and their relationship with tumor angiogenesis and to investigate the interaction of NOS and VEGF in gastric cancer. Methods The expression and distribution of VEGF, iNOS and eNOS in 34 gastric cancer specimens were detected with immunohistochemistry. Microvessel density (MVD) was counted with FⅧRAg immune specific staining. Results The expression rates of iNOS, eNOS and VEGF in 34 gastric cancers were 73.5%, 82.4% and 91.2% respectively. The expression of VEGF had a significant positive relation with iNOS, but not with eNOS. The MVDs of VEGF or iNOS positive gastric cancers were obviously higher than those of VEGF or iNOS negative gastric cancers. There was no significant difference between the MVDs of eNOS positive gastric cancers and eNOS negative ones. Conclusion MVD increases with increase of expression of VEGF and iNOS in gastric cancer. It is indicated that VEGF and iNOS can promote gastric cancer angiogenesis. VEGF and iNOS have a significant positive correlation, which suggests that in human gastric cancer, iNOS plays an important role in the production and action of VEGF.
This study was designed to define the microvessel density (MVD) in breast carcinoma and benign breast disease and the relationship of microvessel density with the tumor size, histologic grade, and lymph node status. Under light microscopy, the microvessels by staining their endothelial cells immunocytochemically for factor Ⅷ were highlighted. Results: The mean level of MVD of breast carcinoma was significantly higher than that of benign disease (Ｐ＜0.01); the MVD of breast carcinoma was associated with tumor size (Ｐ＜0.05), histologic grade (Ｐ＜0.05), and axillary node status (Ｐ＜0.05), but no association with estrogen receptor. These show that MVD of breast carcinoma is significantly higher than that of benign breast disease, and MVD of breast carcinoma is one of significant prognostic indicators.
Objective To explore the effect of toremifene on estrogen receptor （ER） expression and tumor micro-angiogenesis in rat Lewis lung carcinoma. Methods Cell suspension of rat Lewis lung carcinoma was implanted into 40 female Wistar rats subcutaneously. The rats were randomly divided into a control group，a estradiol group （0.006 mg/mL），a low dose toremifene group （0.25 mg/mL） and a high dose toremifene group （5 mg/mL）. Tumor size was measured every 3 days and the tumor growth curve was charted. On 15th day，the tumor weight and the growth inhibition rate were measured. Immunohistochemical method was used to detect the expressions of estrogen receptor α （ERα），estrogen receptor β （ERβ），vascular endothelial growth factor （VEGF），and platelet endothelial cell adhesion molecule-1 （PECAM-1）. Integral optical density （IOD） of ERα，ERβ and VEGF was calculated by image analysis software. Quantitative method of Weidner with PECAM-1 was employed for microvessel density （MVD） count. Results Tumor size of the four groups all presented a quadratic function growth trend with time （Plt;0.05）. Tumor growth speed was slower in toremifene groups of low and high doses than that in the control group and the estradiol group. The growth inhibition rate of the estradiol group，the low dose toremifene group and the high dose toremifene group was －15.1%，22.6%，and 45.1%，respectively. The expressions of ERα，VEGF，and MVD in the estradiol group were significantly higher than those in the control group，the low dose toremifene group and the high dose toremifene group （all Plt;0.05）. The expressions of ERα，VEGF，and MVD in the low dose toremifene group were significantly lower than those in control group，but higher than those in high dose toremifene group （all Plt;0.05）.The expression of ERα was positively related to VEGF （r=0.664，Plt;0.05） and MVD（r=0.593，Plt;0.05）. Conclusion Toremifene can inhibit tumor growth，which maybe involved in inhibiting ERα mediated VEGF expression.
ObjectiveTo investigate the clinicopathological significance of integrin α5β1 expression and microvessel density(MVD) in gastric cancer(GC) and the correlation of MVD with integrin α5β1. MethodsThe expression of integrin α5β1 was detected by means of immunohistochemical staining (SP method) on paraffinembeded tissue specimens from 35 primary gastric carcinoma(PGC), 10 metastasic lymph node of gastric cancer and 8 chronic superficial gastritis (CSG). Vascular endothelial cells were stained immunohistochemically using antiCD34 monoclonal antibody to recognize microvessel(MV) in 35 cases of PGC and 8 CSG, MV was counted in 4 hot spot per slide under lightmicroscope (×400) and the average was defined as MVD. The results combined with clinicopathological parameters were analyzed statistically to characterize the role of integrin α5β1 and MVD in the progression of gastric cancer. ResultsIntegrin α5β1 expression and MVD in PGC were significantly higher than those in CSG respectively （t=3.32, P lt;0.01; t=2.30, Plt;0.05）; the expression of integrin α5β1 in PGC showed only a correlation with the invasion depth of tumor (t=2.29, Plt;0.05） while MVD showed all correlations with invasion depth,lymph node status and TNM stage （t=3.07, Plt;0.01; t=2.48, Plt;0.05; t=2.94,Plt;0.01）. Neither integrin α5β1expression nor MVD showed a relation with differential of PGC （t=0.15, Pgt;0.05; t=0.41, Pgt;0.05）. Integrin α5β1 was significantly overexpressed in lymph node metastatic cancer compared with that in corresponding PGC （t=2.45, Plt;0.05）; the difference of MVD showed no statistical significance among levels of integrin α5β1 expression in PGC （F =1.43,P＞0.05） and it showed no correlation with integrin α5β1 expression（r= 0.156, P=0.37）.Conclusion Overexpression of integrin α5β1 is present in GC and associates with the progression of tumor, implying that it may be viewed as the indicator of invasion and metastasis and the candidate target of gene therapy of gastric cancer. However, integrin α5β1 may not play an important role in the vascularization of GC.
ObjectiveTo explore the expressions of galectin-3 protein and CD105 protein in colorectal cancer and the relationship with clinicopathologic features. MethodsThe expressions of galectin-3 protein and CD105 protein 〔microvessel density (MVD)〕 were detected in 60 cases of colorectal cancer tissues, 30 cases of adenoma tissues, and 30 cases of normal mucosa tissues (at least 4 cm far from carcinoma) by MicrowaveEliVisionTM immunohistochemistry, and the relationship with clinicopathologic features was analyzed. ResultsThe expressions of galectin3 protein and MVD in normal mucosa tissues, adenoma tissues, and cancer tissues gradually increased (Plt;0.05). The expression of galectin-3 protein and MVD in colorectal cancer tissues were correlated to TNM stage, invasive depth, and lymph node metastasis (Plt;0.05， Plt;0.01), and the expression of glectin-3 protein was also correlated to differentiated degree (Plt;0.05). The expression of galectin-3 protein in colorectal cancer tissues was positively correlated to MVD (r=0.420， Plt;0.01). ConclusionsThe high expressions of galectin-3 protein and CD105 protein are correlated to the high invasion ability and lymph node metastasis, which may be potential sensitive index to predict the invasion and metastasis of colorectal cancer.
Objective To investigate the effects of sustained-release basic fibroblast growth factor (bFGF) on healing of bile duct defect. Methods A model of bile duct wall defect (2 cm in length and 1/3-2/3 of the bile duct circumference in width) was made in 24 pigs (male or female, weighing 15-30 kg), and then defect was repaired with sustained-release bFGF collagen membrane (2.0 cm × 1.0 cm × 0.5 cm in size) in the experimental group (n=12) or with collagen membrane (2.0 cm × 1.0 cm × 0.5 cm in size) alone in the blank control group (n=12). Another 4 healthy pigs were used to obtain normal bile duct as normal control group. The survival condition of pigs was observed after operation; at 1, 2, and 3 months after operation, the blood sampling was collected to test the changes of liver function, and the bile duct specimens were harvested to count the microvessel density (MVD) and submucosal gland by HE staining and immunohistochemistry staining; and at 3 months after operation, cholangiography examination was done. Results All the animals survived to completion of the experiment. Intra-abdominal adhesion was serious in the experimental and blank control groups at 1 week after operation, but the adhesion was markedly improved in the experimental group when compared with the blank control group with time passing. The liver function test showed that alkaline phosphatase in the experimental group was significantly lower than that in the blank control group at 2 and 3 months (P lt; 0.05), but no significant difference in aspartate aminortransferase, total bilirubin, and albumin was found among 3 groups (P gt; 0.05). The histology and immunohistochemistry staining observations showed that the regeneration rates of submucosal glands and epithelium in the experimental group were faster than those in the blank control group; defect was covered with the epithelium at 2 months, and the structure was similar to that of normal control group at 3 months; and the edema and inflammation infiltration were reduced when compared with the blank control group. The counts of MVD and submucosal gland were significantly higher than those in blank control group and normal control group at 1 month after operation (P lt; 0.05), and then decreased and remained at normal levels at 2 months after operation. There was a positive correlation between submucosal gland counting and MVD counting in 3 groups after operation (P lt; 0.01). The cholangiography examination showed no biliary dilatation or cholelithiasis after 3 months in experimental group and blank control group. Conclusion Sustained-release bFGF can promote healing of bile duct defect by accelerating the vascularization, gland regeneration, and epithelialization.