Clinicopathological Significance of Tumor Lymphatic Vessel Density in Head and Neck Squamous Cell Carcinoma

Various studies have demonstrated that the lymphatic system is the additional route for solid tumor metastasis. Lymph nodes metastasis in Head and neck squamous cell carcinoma (HNSCC) is a major prognostic indicator for disease progression and a guide for therapeutic strategies. We conducted a study to compare intratumoral (IT) and peritumoral (PT) lymphatic vessel density (LVD) in HNSCC using lymphatic marker D2-40 and its correlation with lymph node metastasis, histological grading and other clinicopathological parameters. Fifty specimen of HNSCC with modified radical neck dissection tissue were included in the study group. Tissue from tumor, peritumoral tissue, tumor margin and all the lymph nodes were processed for paraffin wax blocks and histopathological diagnosis. Immunohistochemical profile of lymphatic vessels in intratumoral and peritumoral tissue was assessed by subjecting one section each from the tumor and peritumoral tissue to D2-40 immunostain. To determine LVD, four fields with the highest LVD (hot spots) were identified. The mean values were calculated by taking an average of all the measurements. The comparison of LVD between peritumoral and intratumoral area revealed significantly higher PT-LVD (P = 0.001). No significant association was seen between LVD, IT-LVD and PT-LVD and different age groups, gender, site of tumor, risk factors, size of tumor, tumor inflammation, pushing/infiltrating margin and stage of tumors. Significantly higher LVD, IT-LVD and PT-LVD was seen in association with lymph node metastasis. Both high intratumoral and peritumoral LVD were found significantly associated with the presence of lymph node metastasis, however lymphatic vessels were found to be significantly more numerous and larger in peritumoral areas as compared to intratumoral lymphatics. The specificity of D2-40 as a lymphatic endothelial marker was also confirmed. The results of our study support the possibility of using the determination of tumor lymphangiogenesis to identify patients of HNSCC who are at risk of developing the lymph node metastasis.     

Differing Lymphatic Vessels Density in Salivary Adenoid Cystic Carcinoma and Pleomorphic Adenoma

Benign and malignant tumours are known to express various factors inducing lymphangiogenesis. Despite their different biological behaviour, salivary pleomorphic adenomas (PA) and adenoid cystic carcinomas (SACC) show similar lymphatic network. Authors compare density of lymphatic network in these tumours. The retrospective study included 20 SACC and 20 PA from salivary tumours. Lymphatic vessel density (LVD) was identified using D2-40 antibody and counted. In SACC, intratumoral, respectively peritumoral, lymphatic vessels were identified in 100 %, respectively 93.8 %, of cases. The intratumoral and peritumoral LVD did not significantly differ from each other. However, they both were higher than normal parenchyma density. In PA, intratumoral LVD, with a single exception, revealed values of 0 and 1. The intratumoral was found to be lower than peritumoral density. The LVD in healthy gland, similar to peritumoral one, was significantly higher than intratumoral values. Direct comparison showed intratumoral and peritumoral LVD in PA to be lower than in SACC. This study comparing LVD in PA and SACC revealed higher values in SACC, outnumbering those in healthy salivary parenchyma and PA. It suggests the capability of this biologically aggressive neoplasm to induce lymphangiogenesis.     

ET-1和VEGF-C在喉癌中的表达及临床意义

目的:研究血管内皮素-1(ET-1)和血管内皮生长因子-C(VEGF-C)在喉癌组织中的表达规律,探讨两者与喉癌发生、发展和淋巴结转移的关系及在预后中的意义。方法:选择45例经病理确诊的喉癌组织为实验组,20例喉良性病变组织为对照组,免疫组化法和实时荧光定量PCR法检测ET-1和VEGF-C蛋白和mRNA的表达,5’-核苷酸酶染色法(5’-Nase)计数淋巴管密度(LVD),CD34染色计数微血管密度(MVD),并结合临床病理特征和生存资料进行相关分析。结果:喉癌淋巴结转移组ET-1、VEGF-C蛋白和mRNA表达显著高于未转移组(P<0.05),二者均显著高于良性喉组织(P<0.01)。喉癌组织中ET-1蛋白表达与瘤内和瘤周LVD、MVD、淋巴结转移、淋巴管浸润、TNM临床分期显著相关(P<0.05),与年龄、性别、肿瘤部位及组织学分级无关(P>0.05)。VEGF-C蛋白表达与瘤内和瘤周LVD、淋巴结转移、淋巴管浸润、MVD显著相关(P<0.05),与年龄、性别、肿瘤部位、TNM临床分期及组织学分级无关(P>0.05)。ET-1和VEGF-C在肿瘤组织中的蛋白表达正相关(r=0.456,P=0.001)。生存分析显示ET-1蛋白阳性表达与生存率无关(P>0.05),ET-1+/VEGF-C+、VEGF-C蛋白阳性表达与生存率负相关(P<0.05),其中ET-1+/VEGF-C+阳性表达更具有显著高危死亡率(P=0.000)。Cox回归模型显示ET-1+/VEGF-C+可以独立影响预后(P<0.05)。结论:ET-1和VEGF-C均能促进喉癌淋巴管生成和血管生成。在喉癌组织中ET-1过表达可能通过诱导VEGF-C表达上调促进喉癌淋巴管生成和淋巴结转移。联合检测ET-1及VEGF-C的表达可成为判断喉癌预后的新的生物学指标。     

胃癌淋巴管密度、微血管密度水平及其临床意义

目的 探讨胃癌组织中淋巴管密度（LVD）和微血管密度（MVD）水平及两者与胃癌临床病理特征的关系。方法 收集本院2004年2月至2007年2月手术切除的68例胃癌患者肿瘤组织,分别采用D2-40和CD31标记胃癌组织中淋巴管和血管,采用免疫组化法检测瘤周及瘤内D2 40、CD31表达情况并计算LVD和MVD,分析瘤周及瘤内LVD、MVD水平与临床病理参数（肿瘤大小、淋巴结转移、脏器转移、TNM分期、性别、年龄、分化程度、Lauren分型及病理类型）和预后的关系。结果 68例患者瘤内和瘤周LVD分别为（4.6±2.0）个和（8.2±3.5）个,瘤内和瘤周MVD分别为（46.3±16.5）个和（47.6±15.3）个;瘤周LVD与肿瘤大小、淋巴结转移、脏器转移及TNM分期有关（P＜0.05）,与性别、年龄、分化程度、Lauren分型及病理类型均无关（P＞0.05）;瘤内LVD、瘤内及瘤周MVD与以上参数均无关（P＞0.05）;全组中位总生存期（OS）为48.6个月,其中瘤周低LVD者的中位OS为31.0个月,低于瘤周高LVD的43.0个月（P＜0.05）;瘤周高MVD者的中位OS为46.0个月,而瘤周低MVD者为48.0个月,差异无统计学意义（P＞0.05）。结论 瘤周LVD与胃癌的临床病理特征及预后密切相关,可能成为胃癌发展及预后的预测指标。     

Peritumoral lymphatic vessel density and vascular endothelial growth factor C expression in early-stage squamous cell carcinoma of the uterine cervix.

PURPOSE: Lymphatic invasion and nodal metastasis plays a major role in the spread of cervical cancer; however, little is known about the mechanisms whereby tumor cells enter the lymphatic system. EXPERIMENTAL DESIGN: We examined the intra- and peritumoral lymphatic vessel density (LVD) using D2-40 immunohistochemistry in 111 cervical squamous cell carcinomas and correlated them with vascular endothelial growth factor (VEGF)-C expression, clinicopathologic tumor features, and outcome. RESULTS: Compared with benign cervix, intratumoral and peritumoral LVD was significantly increased (P < 0.0001). Peritumoral LVD was significantly higher than intratumoral LVD (P = 0.009). High peritumoral, but not intratumoral, LVD showed significant correlation with high tumor stage, lymphatic invasion, and nodal metastasis. VEGF-C showed increased expression at the invasive edge compared with the center of tumors (P < 0.0001) and correlated with high peritumoral LVD, lymphatic invasion, and nodal metastasis. High peritumoral LVD and VEGF-C expression at the invasive edge of tumors were associated with poor overall and recurrence-free survival in univariate analysis. In multivariate analysis, peritumoral LVD was the only independent term predictive of overall survival. CONCLUSIONS: Our findings suggest a potential role for VEGF-C in tumor-induced lymphangiogenesis represented by high peritumoral LVD, which may be one of the mechanisms leading to lymphatic invasion and metastatic spread. High peritumoral LVD may be an independent prognostic factor in early-stage cervical cancer.     

直肠良恶性肿瘤中肿瘤相关巨噬细胞和微血管及淋巴管生成的关系

目的:探讨直肠良恶性肿瘤组织中微血管密度(MVD)及淋巴管密度(LVD)与肿瘤相关巨噬细胞(TAMs)数量的相关性。方法:采用免疫组化的方法,用CD68标记TAMs,分别用CD31和D2-40标记微血管和淋巴管,光镜下对TAMs,MVD及LVD进行计数,分析。结果:直肠腺癌组织中的TAMs,MVD及LVD计数明显高于直肠腺瘤型息肉组织(P<0.01);在直肠腺癌组中,TAMs与MVD和LVD有明显相关性(TAMs与MVD:P<0.01;TAMs与LVD:P<0.01),且MVD与直肠癌淋巴结转移(P<0.01)和分化程度(P<0.05)密切相关;而TAMs和LVD仅与淋巴结转移有显著相关性(P<0.01,P<0.05)。结论:TAMs,MVD及LVD可能是反映直肠腺癌发生、发展、转移及侵袭能力的生物学指标,TAMs在肿瘤中的浸润可能与肿瘤血管及淋巴管的生成有关。     

乳腺癌患者新辅助化疗前后淋巴管密度检测

背景与目的:淋巴管生成在肿瘤经淋巴途径转移中所起的作用已成为近几年里新的研究热点,但目前的研究结果仍存在很多争议。本研究拟检测乳腺癌患者新辅助化疗前后的淋巴管密度(lymphatic vessel density,LVD)并与乳腺良性病变进行比较,以进一步明确淋巴管生成的意义及其在化疗过程中的变化。方法:45例原发性乳腺癌患者给予3个周期新辅助化疗并手术,免疫组化法标记肿瘤及26例纤维腺瘤标本中特异性淋巴管内皮标志物Podop lan in并计数LVD。结果:乳腺癌瘤周、瘤内的淋巴管阳性率分别为70.9%、19.8%。化疗前瘤周平均LVD为9.6,纤维腺瘤瘤周平均LVD为5.4,两者间差异有显著性(P=0.012)。化疗后瘤周平均LVD为3.3,与化疗前相比显著降低(P<0.001),化疗前、后瘤内平均LVD分别为1.9、0.9(P=0.055)。淋巴结转移阳性和阴性组的平均瘤周LVD分别为11.8、7.0(P=0.048)。化疗后客观缓解组化疗前后平均瘤周LVD值分别为9.8、2.0(P<0.001)。结论:与良性病变相比,乳腺癌瘤周LVD显著增高;有瘤内淋巴管存在;瘤周LVD与HER-2、腋淋巴结转移状态相关;化疗后瘤周LVD明显降低,而瘤内LVD无显著变化;化疗后瘤周LVD的降低程度与化疗疗效相关。     

Expression of Vascular Endothelial Growth Factor-C and Vascular Endothelial Growth Factor Receptor-3 in Ovarian Epithelial Tumors

Objective： To explore the role of vascular endothelial growth factor-C （VEGF-C） in the process of angiogenesis, lymphangiogenesis and lymphatic metastasis in epithelial ovarian tumors. Methods： In situ hybridization and immunohistochemical staining for VEGF-C were performed in 30 epithelial ovarian carcinomas, 9 borderline tumors and 26 benign tumors. Endothelial cells were immunostained with anti-VEGFR-3 pAb and anti-CD31 mAb, and VEGFR-3 positive vessels and microvessel density （MVD） were assessed by image analysis. Results： VEGF-C mRNA and protein expression were detected in cytoplasm of carcinoma cells. VEGF-C mRNA and protein expression in ovarian epithelial carcinomas were significantly higher than those in borderline tumors and benign tumors （P〈0.05 or P〈0.01）. In ovarian epithelial carcinomas, VEGF-C protein expression, VEGFR-3 positive vessels and MVD were significantly higher in the cases of clinical stage Ⅲ-Ⅳ and with lymph node metastasis than those of clinical stage Ⅰ-Ⅱ and without lymph node metastasis respectively （P〈0.05 or P〈0.01）. VEGFR-3 positive vessels and MVD were significantly higher in VEGF-C protein positive tumors than negative tumors （P〈0.05）. VEGFR-3 positive vessels was significantly correlated with MVD（P〈0.01）. Conclusion： VEGF-C might play a role in lymphatic metastasis via lymphangiogenesis and angiogenesis in epithelial ovarian tumors, and VBEGF-C could be used as a biologic marker of metastasis in ovarian epithelial tumors.     

VEGF-C 及VEGFR-3 在肝细胞癌中的表达

 目的　探讨VEGF-C、VEGFR-3、微血管密度（MVD）、微淋巴管密度（LVD）及肝细胞癌临床病理特征之间的关系。方法　取肝细胞癌组织标本60例，正常肝组织标本20例。采用反转录聚合酶链反应（RT-PCR）方法分析其中VEGF-C及VEGFR-3 mRNA 的表达，以免疫组织化学法检测肝癌MVD及LVD，并分析四者与肝癌临床病理特点之间的关系。结果　肝癌组织VEGF-C、VEGFR-3 mRNA表达、MVD及LVD高于正常肝组织（P＜0.01）；肝癌组织中，VEGF-C与VEGFR-3表达、MVD及LVD均呈正相关（P＜0.01），VEGF-C及VEGFR-3表达与肝癌肝内转移、门静脉癌栓形成及淋巴转移相关（P＜0.01），MVD与肝癌肝内转移、门静脉癌栓形成相关（P＜0.01），LVD与淋巴转移相关（P＜0.01）。结论　肝细胞癌组织中VEGF-C及VEGFR-3表达增多，可能通过参与血管、淋巴管生成促进肿瘤的侵袭、转移。     

Clinicopathological significance of peritumoral lymphatic vessel density in gastric carcinoma

Lymphangiogenesis has recently been considered important for spread of malignant tumors. In the present study, lymphatic vessel density (LVD) including peritumoral LVD (P-LVD) and intratumoral LVD (I-LVD) was determined, respectively, by immunohistochemical staining with the antibody to LYVE-1 in 63 cases of early gastric carcinoma and 105 cases of advanced gastric carcinoma. The aim of the study is to investigate whether or not increased LVD could be a risk factor for nodal metastasis and survival. We conclude that increased P-LVD, but not I-LVD, could serve as an independent risk factor for nodal metastasis, recurrence and overall survival in gastric carcinoma.     

MIA-dependent angiogenesis and lymphangiogenesis are closely associated with progression,nodal metastasis and poor prognosis in tongue squamous cell carcinoma

We examined the role of angiogenesis/lymphangiogenesis and the relationship between melanoma inhibitory activity (MIA) and angiogenesis or lymphangiogenesis in oral squamous cell carcinoma (OSCC). One hundred and one formalin-fixed, paraffin-embedded specimens of primary OSCC were evaluated for microvessel density (MVD), lymphovessel density (LVD), expression of vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D and MIA. Fresh frozen 18 samples of primary OSCC were further examined for the expression of VEGF, VEGF-C, VEGF-D and MIA protein by enzyme-linked immunosorbent assay (ELISA). In in vitro analysis, we studied the change of VEGF, VEGF-C and VEGF-D expression after MIA siRNA treatment. Higher MVD, LVD and VEGF expression levels were closely associated with tumour progression, nodal metastasis and poor prognosis. Expression levels of VEGF-C and VEGF-D were only related with nodal metastasis. MIA expression was significantly associated with MVD, LVD, VEGF, VEGF-C and VEGF-D expression by immunohistochemistry and ELISA assay. VEGF, VEGF-C, VEGF-D and MIA expression levels of metastatic tongue cancer HSC-3 cells were higher than those with no metastatic HSC-4 cells, and VEGF, VEGF-C and VEGF-D expression levels were decreased by MIA siRNA treatment in both cells. MIA-dependent angiogenesis/lymphangiogenesis might be a useful therapeutic target in progressive and metastatic OSCC.     

Carcinoma of salivary glands--a clinical analysis of 342 cases

342 cases with carcinoma of salivary glands were analysed. 152 tumors were located in parotid, 42 in submaxillary, 17 in sublingual and 131 in minor salivary glands. Pathological diagnosis were 106 mucoepidermoid carcinoma, 80 adenoid cystic carcinoma, 54 malignant mixed tumor, 40 adenocarcinoma, 38 papillary cystadenocarcinoma, 17 acinic cell carcinoma, 5 squamous cell carcinoma, and 2 undifferentiated carcinoma. The 3, 5, 10 and 15 year survival rates of these 342 cases were 76.6%, 65.9%, 48% and 29%, respectively. The difference between survival rate and relapse-free survival rate was about 8%. The prognosis of acinic cell carcinoma and mucoepidermoid carcinoma was much better than that of squamous cell carcinoma, adenocarcinoma and undifferentiated carcinoma. The survival rates, according to location, were: minor salivary gland tumors the highest, and those of submaxillary gland tumors the lowest. Postoperative radiotherapy improved the survival rate of adenoid cystic carcinoma. The overall recurrence rate was 37.4%, the neck lymph node metastasis rate 14.3% and the distant metastasis rate 9.1%.     

Angiogenesis and Lymphangiogenesis in Oral Squamous Cell Carcinoma: Comparison of Japanese and Indian Cases

A comparative study between 17 Japanese and 19 Indian patients with oral squamous cell carcinomas (OSCCs)revealed that the tumour prognostic indicator mean vessel density (MVD) count for angiogenesis was relativelyhigh at 57.1 in Indian as compared to 39.3 in Japanese (P=0.001) cases, whereas the lymph-vessel density (LVD)count for lymphangiogenesis was lower (12.8 vs 48.0, P=0.002). Both male and female Indians had higher MVDcounts, but LVD counts were only slightly lower in females. MVD count was relatively high among the casesbelow 65 years old in both the countries (P=0.4). Japanese cases with Tongue cancer had higher MVD count,but the Indian cases had lower LVD counts. Size-wise, T2 and T3 had higher counts of MVD both in Indian andJapanese cases. MVD and LVD count was higher in grades II and III both in Japanese and Indian cases. Therewas insignificant difference of the MVD counts among smokers, but the tobacco chewers in Indian cases hadhigher counts of MVD and LVD (P value by Bartlett test 0.35, 0.57 respectively). The hot-spots of tumour siteshad variable rates of lymphocyte infiltration showed higher MVD counts in all the cases. Although the clinicalcharacteristics and demographic variables usually relate to MVD and LVD counts, the tendency of higher values,especially among tobacco chewers, identified as the highest risk group for occurrence of oral cancer needs to beinvestigated further.     

Multi-kinase inhibitor E7080 suppresses lymph node and lung metastases of human mammary breast tumor MDA-MB-231 via inhibition of vascular endothelial growth factor-receptor (VEGF-R) 2 and VEGF-R3 kinase

PURPOSE: Vascular endothelial growth factor (VEGF)-C/VEGF-receptor 3 (VEGF-R3) signal plays a significant role in lymphangiogenesis and tumor metastasis based on its effects on lymphatic vessels. However, little is known about the effect of inhibiting VEGF-R3 on lymphangiogenesis and lymph node metastases using a small-molecule kinase inhibitor. EXPERIMENTAL DESIGN: We evaluated the effect of E7080, a potent inhibitor of both VEGF-R2 and VEGF-R3 kinase, and bevacizumab on lymphangiogenesis and angiogenesis in a mammary fat pad xenograft model of human breast cancer using MDA-MB-231 cells that express excessive amounts of VEGF-C. Lymphangiogenesis was determined by lymphatic vessel density (LVD) and angiogenesis by microvessel density (MVD). RESULTS: In contrast to MDA-MB-435 cells, which expressed a similar amount of VEGF to MDA-MB-231 cells with an undetectable amount of VEGF-C, only MDA-MB-231 exhibited lymphangiogenesis in the primary tumor. E7080 but not bevacizumab significantly decreased LVD within the MDA-MB-231 tumor. E7080 and bevacizumab decreased MVD in both the MDA-MB-231 and MDA-MB-435 models. E7080 significantly suppressed regional lymph nodes and distant lung metastases of MDA-MB-231, whereas bevacizumab significantly inhibited only lung metastases. E7080 also decreased both MVD and LVD within the metastatic nodules at lymph nodes after resection of the primary tumor. CONCLUSIONS: Inhibition of VEGF-R3 kinase with E7080 effectively decreased LVD within MDA-MB-231 tumors, which express VEGF-C. Simultaneous inhibition of both VEGF-R2 and VEGF-R3 kinases by E7080 may be a promising new strategy to control regional lymph node and distant lung metastases.     

Loss of Maspin expression is a negative prognostic factor in common salivary gland tumors

Maspin, a 42kDa protein, belongs to the serine protease inhibitor (serpin) family and is suggested to have inhibitory effects on tumor-induced angiogenesis, tumor cell motility, invasion and metastasis and influences prognosis of tumor patients. The aim of the study was to analyze Maspin expression in salivary gland cancer as well as its prognostic impact on survival in comparison to clinical parameters. Immunohistochemical staining was carried out in 73 cases of salivary gland malignancies. High proportions of Maspin expression were observed in adenoid cystic carcinomas, mucoepidermoid carcinomas and carcinomas ex pleomorphic adenoma, low proportions were seen in salivary duct carcinomas. Acinic cell carcinomas did not show any Maspin expression. Analysis of the prognostic impact of Maspin expression was restricted to salivary gland carcinoma types of intermediate malignancy grade (adenoid cystic carcinoma, mucoepidermoid carcinoma and carcinoma ex pleomorphic adenoma). For these tumors, univariate analyses revealed that T-stage (p=0.025), age70 (p=0.0065), loss of Maspin (p=0.0016) and presence of residual tumor (p<0.001) correlated with poor prognosis. In multivariate analysis age70 (p=0.005) and loss of Maspin (p=0.036) were significant prognostic factors. Moreover, negative Maspin staining was associated with lymph node metastasis and residual tumor. According to these findings, Maspin might be useful as a new prognostic marker in adenoid cystic carcinoma and in salivary gland carcinomas with intermediate grade of malignancy where grading systems are still under debate.     

淋巴管密度在甲状腺乳头状癌和滤泡癌的表达及意义

[目的]探讨淋巴管密度（LVD）在甲状腺乳头状癌（PTC）和滤泡癌的表达及意义。[方法]采用免疫组化技术,以D2-40标记淋巴管内皮,CD31标记血管内皮,对105例PTC、18例甲状腺滤泡癌和21例滤泡性腺瘤进行LVD和微血管密度（MVD）的检测。对LVD与乳头状癌的临床病理资料的关系,及乳头状癌和滤泡癌中MVD和LVD的表达差异进行了统计分析。[结果]（1）PTC中均可见D2-40阳性淋巴管,其瘤周LVD较瘤内为高,淋巴结转移组（瘤内6.80±4.40;瘤周13.63±5.09）较无转移组（瘤内4.30±3.06;瘤周10.17±4.53）为高,两组比较有统计学差异（P〈0.01）。（2）甲状腺乳头状癌的瘤内LVD与TNM分期有关,高分期组（Ⅲ期/Ⅳ期）的瘤内LVD（7.33±4.79）较低分期组（Ⅰ期/Ⅱ期,5.16±3.71）为高,两者相比有统计学差异（P〈0.05）。（3）甲状腺乳头状癌的瘤内LVD（5.47±3.93）较滤泡癌的瘤内LVD（1.56±1.92）为高,两者相比,差异具有统计学意义（P〈0.05）。[结论]甲状腺乳头状癌LVD与淋巴结转移密切相关,瘤周淋巴管的扩张形态和相对较高的LVD在甲状腺乳头状癌淋巴道转移中可能发挥了重要的作用。     

Tumor lymphangiogenesis correlates with lymph node metastasis and clinicopathologic parameters in oral squamous cell carcinoma

BACKGROUND: Lymphatic vessel density (LVD) and microvessel density (MVD) are important parameters for assessing the malignant potential of tumors and patient survival. In this report, the authors defined LVD as the density of D2-40-positive lymphatic vessels and MVD as the density of CD105-positive microvessels per unit area of tissue. It was reported previously that vascular endothelial growth factor C (VEGF-C) is a major modulator of LVD and MVD. The objectives of this study were to clarify the clinical and prognostic significance of both LVD and MVD in oral squamous cell carcinoma (OSCC) and to elucidate the lymphangiogenic and angiogenic activities of VEGF-C in cancer tissues. METHODS: In total, 110 OSCC tissue samples were evaluated for LVD, MVD, and expression of VEGF-C using immunohistochemistry. Correlations among these parameters and clinicopathologic factors were examined. RESULTS: LVD was significantly higher in tumors that had very high expression of VEGF-C compared with tumors that had no/weak expression of VEGF-C. LVD correlated well with lymph node metastasis (P < .001). MVD was correlated significantly with positive lymph node metastasis (P < .001) but not with VEGF-C expression. In contrast, high expression of VEGF-C was correlated significantly with advanced tumor status (P = .041). Survival rates were lower in patients who had higher LVD (P < .001), higher MVD (P = .0028), and strong VEGF-C expression (P = .048). CONCLUSIONS: Lymphangiogenesis predominantly influenced metastasis-free survival. The current results suggested that LVD is a more useful tool than MVD and VEGF-C for deciding on therapeutic strategies in patients with OSCC.     

Expression of vascular endothelial growth factors-C and -D correlate with evidence of lymphangiogenesis and angiogenesis in pancreatic adenocarcinoma

Background: We analyzed the intratumoral and peritumoral microvessel density (MVD) and microlymphatic vessel density (MLVD) in pancreatic adenocarcinoma (PAC) and recorded the expression of vascular endothelial growth factor (VEGF)-C and -D. These data were tested for their significance for tumor progression. Methods: The tissue samples were obtained from 30 patients with PAC. The expression of VEGF-C and -D, MLVD, MVD was assayed by immunohistochemical staining. The expression of VEGF-A and -C, and -D mRNA was detected by semi-quantitative RT-PCR. Results: Immunohistochemical analysis revealed the presence of VEGF-C and -D immunoreactivity in 73% (22/30) and 57% (17/30). The positive rates of VEGF-C and -D protein in central portion of tumors (30% and 16.7%) were significantly lower than those in marginal portion (73.3% and 56.7%). The group with high expression of VEGF-C and -D in marginal portion had higher incidence of lymph node metastasis, lymphatic invasion and venous invasion. The MLVD in both of the VEGF-C and -D positive groups was higher than that in the negative groups, and the lymph node metastasis increased. MVD in the VEGF-C positive group was higher than that in the negative group. Conclusions: The expression of VEGF-C and -D in the marginal portion of tumor significantly associated with lymphatic metastasis and prognosis in patients with PAC, and may induced lymphangiogenesis. VEGF-C was important in the regulation of angiogenesis and lymphangiogenesis in PAC.     

人肝癌组织中iNOS、VEGF的表达及微血管密度的病理意义

背景与目的:诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和血管内皮生长因子(vascular endothelial growth factor,VEGF)被认为是诱导和调节肿瘤血管生成,进而影响肿瘤病理进展和预后的重要相关因子。本研究检测人肝细胞癌(hepatocellular carcinoma,HCC)及相应癌旁组织中iNOS、VEGF的表达,探讨其与血管生成的关系,为临床诊治HCC及判断其预后提供理论依据。方法:应用组织芯片技术,采用原位杂交和免疫组化法分析147例HCC组织及癌旁组织中iNOS、VEGF的表达,并用CD34标记免疫组化法检测微血管密度(microvessel density,MVD)。结果:iNOS、VEGF在癌旁组织中的阳性率分别为33.33%和40.82%,而在癌组织中的阳性率分别为86.39%和78.91%,癌组织与癌旁组织比较差异有显著性(P<0.01)。癌组织的MVD值为56.5±12.8,癌旁组织的MVD值为8.4±3.6,两者差异有显著性(P<0.01)。iNOS的表达与肿瘤大小、乙型肝炎表面抗原(HBsAg)相关(P<0.05),而与转移和肿瘤分化程度无关(P>0.05)。VEGF的表达及MVD值与肿瘤大小、转移相关(P<0.05),而与HBsAg和肿瘤分化程度无关(P>0.05)。在癌组织中MVD与VEGF、iNOS的表达呈正相关,VEGF和iNOS之间亦存在正相关关系(P<0.01)。结论:HCC中iNOS及VEGF的表达与肿瘤血管生成有关。癌组     

Expressions of COX-2 and VEGF-C in gastric cancer: correlations with lymphangiogenesis and prognostic implications

Background

Cyclooxygenase-2 (COX-2) has recently been considered to promote lymphangiogenesis by up-regulating vascular endothelial growth factor-C (VEGF-C) in breast and lung cancer. However, the impact of COX-2 on lymphangiogenesis of gastric cancer remains unclear. This study aims to test the expression of COX-2 and VEGF-C in human gastric cancer, and to analyze the correlation with lymphatic vessel density (LVD), clinicopathologic features and survival prognosis.

Methods

Using immunohistochemistry, COX-2, VEGF-C and level of LVD were analyzed in 56 R0-resected primary gastric adenocarcinomas, while paracancerous normal mucosal tissues were also collected as control from 25 concurrent patients. The relationships among COX-2 and VEGF-C expression, LVD, and clinicopathologic parameters were analyzed. The correlations of COX-2, VEGF-C and level of LVD with patient prognosis were also evaluated by univariate tests and multivariate Cox regression.

Results

The expression rates of COX-2 and VEGF-C were 69.64% and 55.36%, respectively, in gastric carcinoma. Peritumoral LVD was significantly higher than that in both normal and intratumoral tissue (P < 0.05). It was significantly correlated with lymph node metastasis and invasion depth (P = 0.003, P = 0.05). VEGF-C was significantly associated with peritumoral LVD (r = 0.308, P = 0.021). However, COX-2 was not correlated with VEGF-C (r = 0.110, P = 0.419) or LVD (r = 0.042, P = 0.758). Univariate analysis showed that survival time was impaired by higher COX-2 expression and higher peritumoral LVD. Multivariate survival analysis showed that age, COX-2 expression and peritumoral LVD were independent prognostic factors.

Conclusions

Although COX-2 expression was associated with survival time, it was not correlated with VEGF-C and peritumoral LVD. Our data did not show that overexpression of COX-2 promotes tumor lymphangiogenesis through an up-regulation of VEGF-C expression in gastric carcinoma. Age, COX-2 and peritumoral LVD were independent prognostic factors for human gastric carcinoma.