目的 通过复制人肝癌细胞株HepG2裸鼠皮下移植瘤模型，观察绿茶提取物表没食子儿茶素没食子酸酯（EGCG）干预对HepG2移植瘤新生血管生成的影响。 方法 瘤体接种复制HepG2移植瘤模型，荷瘤裸鼠20只随机分组，实验组给予EGCG溶液每日20 mg/（kg·只），腹腔注射3周，对照组给予等量灭菌注射用水3周，末次用药24 h，后处死裸鼠，剥离移植瘤。常规病理切片观察移植瘤组织结构；逆转录-聚合酶链式反应和免疫组织化学法检测移植瘤缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）mRNA及蛋白表达，并通过检测CD34表达计数瘤组织微血管密度（MVD）。 结果 组织病理学观察实验组移植瘤见大量坏死区，瘤体内血管数量明显少于对照组；实验组HIF-1α、VEGF mRNA及蛋白表达水平比对照组均明显下调（P＜0.05），实验组MVD比对照组明显下降（P＜0.05）。 结论 EGCG可抑制荷瘤裸鼠HepG2移植瘤新生血管生成。
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.
摘要:目的: 探讨选择性内皮素A受体拮抗剂BQ123对人喉癌Hep2细胞裸鼠种植瘤的生长及血管形成的影响。 方法 :将实验动物裸鼠随机分为3组:BQ123［n =8,2mg/(kg·day)］、氟尿嘧啶组［n =8,2mg/(kg·day)］、生理盐水组(n =8),比较各组裸鼠成瘤体积、微血管密度(MVD)。 结果 :BQ123组肿瘤体积为(162±053)cm3,明显小于生理盐水组及氟尿嘧啶组,差异具有统计学意义;BQ123组的肿瘤组织中MVD高倍镜下为232,明显低于生理盐水组(586)及氟尿嘧啶组(395),差异具有统计学意义。 结论 :BQ123对人喉癌Hep2细胞在裸鼠体内有明显抑瘤作用,肿瘤的体积、肿瘤组织MVD显著低于对照组,表明BQ123可通过抑制肿瘤血管生成而显著抑制肿瘤生长。Abstract: Objective: To study the effects of endothelin A receptor blockade BQ123 on the implanted human laryngeal carcinoma angiogenesis of nude mouse. Methods : From March 2008 to July 2009, 24 Balb/c nude mice were randomly divided into three groups: BQ123 group ［〖WTBX〗n =8, BQ123 at 2mg/(kg·day)］, 5Fu group ［〖WTBX〗n =8, fluorouracil at 2mg/(kg·day)］ and the control group (〖WTBX〗n =8, normal saline). The carcinoma volume and microvascular density of each group were compared. Results : The tumor size of BQ123 group, which was (162±053)cm3 in average, was significant smaller than the tumor sizes of the other two group s. The average microvascular density score of the tumors in BQ123 group was 232 per hyper power len (HP), which was also significantly less than the average scores of control groups (586 and 395 respectively). Conclusion : Nude mouse experiments show that the carcinoma volume and microvascular density of BQ123 group are significantly lower than those of the control groups. BQ123 inhibits the growth of carcinoma by its inhibition of carcinoma angiogenesis.
Objective To investigate the enhancement of the transverse rectusabdominis musculocutaneous (TRAM) flap survival in local ischemic area by recombinant adenovirus mediated vascular endothelial growth factor 165 gene(Ad-VEGF-165). Methods The vascular pedicle TRAM flaps were made in the right abdomin of30 SD rats and they were randomly divided into 5 groups. The Ad-VEGF-165 was injected into the subcutaneous tissue of epigastra(group 1), the subcutaneous tissue of epigastria and rectus abdominis muscle (group 2), and the rectus abdominis muscle(group 3); Adenovirus mediated green fluorescent protein(Ad-GFP) and DMEMwere injected into the subcutaneous tissue of epigastria and rectus abdominis muscle as autocontrol(group 4) and blank control(group 5), respectively. The survival areas of TRAM flap was measured after 7 days of operation. The microvascular density(MVD) and the integral optical density (IOD) were tested with anti-rat CD34 and with VEGF immunohistochemistry and insitu hybridization histochemistry (ISHH), respectively. Results The survivalareas of TRAM flap in groups 1, 2 and 3 (14.19±2.77, 15.18±2.18 and 8.30±1.28 cm2) were higher than those in groups 4 and 5(4.12±186 and 3.60±1.95 cm2), being significant differences(Plt;0.05).The CD34 MVD of the TRAMflap in groups 1, 2 and 3 was higher than that in groups 4 and 5; the positiveexpression for VEGF and ISHH were shown in groups 1, 2 and 3 and there was significant difference when compared with groups 4 and 5 (Plt;0.05). Conclusion Treatment by recombinant Ad-VEGF165gene is an effective option for enhancement of the TRAM flap survival in the local ischemic area.
Objective To investigate the relationship of vascular endothelial growth factor (VEGF), microvessel density (MVD) and progression of gastric carcinoma (GC). Methods The expression of VEGF and MVD in archival waxembedded specimens of 80 cases of GC and 20 gastric benign disease (GBD) were examined by using immunohistochemical staining. ResultsThe positive expression rate (PER) of VEGF in GC was 75.0%, and in GBD 5.0% (P＜0.05). The PER of VEGF in GC with invasive serosa was 95.5%, in those without serosal invasion 50.0% (P＜0.05). 82.8% was the PER of VEGF in GC with lymph node metastasis, 54.5% without lymph node metastasis (P＜0.05).The PER of VEGF in GC accompanied by distant metastasis was 100%, higher than that without distant metastasis (71.0%, P＜0.05). PER of VEGF in pTNM Ⅰ+Ⅱ was 53.1%, in Ⅲ+Ⅳ 89.6% (P＜0.05). MVD correlated significantly with depth of invasion, lymph node metastasis,distant metastasis and pTNM stages (P＜0.05). There was correlationship between MVD and VEGF (P＜0.05).Conclusion VEGF expression upregulation and MVD contribute to the progression of gastric carcinoma.
ObjectiveTo study the mechanism of reducing the intratumoral microvessel density (MVD) by Ginsenoside Rg3 (Rg3) combined with cytotoxic agent in xenotransplanted human breast infiltrating duct carcinoma in nude mice. MethodsSixteen female nude mice were randomly divided into 4 groups to receive cyclophosphamid (16 mg/kg,qd) combined with Rg3 (10 mg/kg, qd),Rg3(10 mg/kg,qd) alone,cyclophosphamid (16 mg/kg,qd) alone and 0.5% sodium carboxymethyl cellulose (0.5 ml,qd) respectively for 55 days. Breast cancer mass were weighed and sampled for light microscopic observation. The intratumor MVD was examined by immunohistochemical staining. ResultsThe tumor weight of treated group was significantly lower than that of control group. The tumor weight of the Rg3 combined with CTX group was lower than that of Rg3 group. The MVD value of Rg3 group was significantly lower than that of CTX group and control group. The MVD was significantly reduced in the Rg3 combined with CTX group than that in the others.ConclusionRg3 combined with CTX can inhibit the growth of xenotransplanted human breast infiltrating duct carcinoma, and reduce the intratumoral MVD.
The present paper aims to investigate whether or not vasculogenic mimicry (VM) exists in laryngeal squamous cell carcinoma (LSCC), and to elucidate its relationship to microvessel density (MVD), galectin-3 (Gal-3) expressionb and clinicopathological factors of patients with LSCC. VM, score of MVD and expression of Gal-3 protein were detected by immunohistochemistry and histochemistry in 83 specimens of LSCC tissue and 20 specimens of normal laryngeal tissue. The positive rate of VM in normal laryngeal tissues was 0%, and was 33.7% in LSCC tissues. There was a significant difference between the two groups (P<0.01). VM or MVD was significantly related to differentiation, pTNM stages and lymph node metastasis of LSCC (P<0.05), but not to age, gender and tumor site (P>0.05). And there was a positive correlation between every two of VM, score of MVD, and Gal-3 protein (P<0.05). The results suggest that expression of Gal-3 protein may be related to the initiation, angiogenesis and VM formation in LSCC; And VM, angiogenesis and Gal-3 protein may be involved in the development, invasion and metastasis of LSCC.
Objective To investigate the expression levels and significance of vascular endothel ial growth factor (VEGF) and microvessel density (MVD) in rabbit radius defects repaired with allogeneic and autogenic bone. Methods Forty adult New Zealand rabbits were chosen, and 10 mm bone defect model was created in the bilateral radii of 28 experimental rabbits. The other 12 rabbits provided allogeneic bone under the standard of American Association of Tissue Bank. In the left side, allogeneic bone were used to repair bone defect (experimental group), equal capacity autogenous il iac bone was used in the right side (control group). Animals were sacrificed at 2, 4, 8, and 12 weeks postoperatively. Immunohistochemical method was used to determine the expression of VEGF, CD34 protein and MVD counting. Bone histomorphometric parameters, including percent trabecular area (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp) were measured by von Kossa staining undecalcified sl ices. The relation was analyzed between VEGF and MVD, histomorphometric parameters. Results The positive signals of VEGF protein were detected in cytoplasm of vascular endothel ial cells, chondrocytes, osteoblasts, fibroblasts and osteoclasts. At 2 weeks, there was no significant difference in VEGF protein expression between experimental group and control group (P gt; 0.05); at 4 and 8 weeks, the expression of VEGF in control group was significantly higher than that in experimental group (P lt; 0.05); and at 12 weeks, there was no significant difference between two groups (P gt; 0.05). There was a positive correlation (P lt; 0.01) between VEGF expression and MVD value in two groups at 2, 4, 8, and 12 weeks postoperatively. There was no significant difference in bone histomorphometric parameters (BV/TV, Tb.Th, Tb.N, Tb.Sp) between two groups at 12 weeks postoperatively (P gt; 0.05), but there was a positive correlation between VEGF expression and parameters of BV/TV, Tb.Th, and Tb.N (P lt; 0.01); and a negative correlation between VEGF and Tb.Sp (P lt; 0.01). Conclusion VEGF can express diversity at different time and positions, and the different expressions indicated various biology significances in the process of the bone heal ing. It can coordinate growth of cartilage and bone and profit vascular reconstruction of allogeneic bone. VEGF may participate in promoting osteogenesis in the course of allogeneic bone transplantation.
Objective To investigate perfusion features of gastric antrum cancer by 64-multidetector CT and to assess the correlation between perfusion CT parameters and immunohistochemical markers of angiogenesis in gastric cancer. Methods Perfusion CT was performed in 30 patients with gastric antrum cancer （gastric antrum cancer group） and 24 patients with normal stomach (control group), and postoperative specimens were stained using a polyclonal antibody to VEGF and CD34. The correlation between perfusion parameters and microvessel density (MVD), and VEGF were analyzed. Results Blood volume (BV) increased in the gastric antrum cancer group (Plt;0.01). There was no significant difference in perfusion (PF), peak enhancement (PE), or time to peak (TTP) between the gastric antrum cancer and the normal groups (Pgt;0.05). BV was positively significantly correlated with MVD （r=0.522, P=0.02), but no significant correlation was found between PF （r=0.072, P=0.78), PE （r=0.253, P=0.31), or TTP （r=0.235, P=0.35) and MVD. No correlation was found between PF （r=-0.208, P=0.45), PE （r=-0.251, P=0.37), TTP（r=-0.284, P=0.31), or BV（r=-0.472, P=0.09) and VEGF.Conclusion Blood volume can evaluate the angiogenesis of tumor and perfusion CT can be a tool to assess microvessel status in gastric antrum cancer.