High Lymphatic Microvessel Density Research Articles

A meta-analysis of the lymphatic microvessel density and survival in gastric cancer with 1809 cases.

Lymph node metastasis commonly occurs in gastric cancer. Previous studies have demonstrated that the overexpression of lymphatic microvessel density (LVD) is correlated with various malignancies. To evaluate the potential role of LVD in various malignancies, we conducted a systematic review and meta-analysis to thoroughly investigate the association of LVD expression with tumor progression and survival in gastric cancer. We performed a comprehensive search of common databases and selected studies demonstrating the relationship between LVD expression and gastric cancer prognosis. Hazard ratios (HR) were used to determine the value of LVD for predicting gastric cancer metastasis and prognosis. The data were extracted from the included studies and pooled with the appropriate effects model using STATA 12.0. The results showed that high LVD expression obviously impacted the prognosis of gastric cancer, based on an overall survival (OS) HR of 2.58 (95% CI: 1.91–3.48, P < 0.001) and a disease-free survival (DFS) HR of 2.51 (95% CI: 1.35–4.68, P = 0.004) in the univariate analysis. In addition, the results of the multivariate analysis indicated a remarkable relationship between high LVD expression and gastric neoplasm prognosis. The pooled OS HR was 4.12 (95% CI: 3.45–4.91, P < 0.001). The current meta-analysis shows that high LVD is closely related to tumor metastasis and poor prognosis in gastric malignancy. LVD could be a key factor in tumor lymphatic metastasis. Moreover, LVD is likely a potential index and an effective biomarker for the prediction of patient prognosis.

Lymphangiogenesis and colorectal cancer.

Lymphatic metastasis is an important event in the progress of metastasis in colorectal cancer (CRC). The purpose of this article is to assess the role of lymphangiogenesis on CRC. In peritumoral areas of CRC, the lymphatic microvessel density (LMVD) is higher than those in normal colorectal tissues. Morever, the high LMVD is correlated with DFS and local recurrence in CRC. The VEGF-C/VEGF-D/VEGFR-3 pathway, sonic hedgehog (Shh) signaling pathway and extracellular matrix (ECM) are involved in the regulation of lymphangiogenesis in CRC. Inhibition of the VEGF-C/VEGF-D/VEGFR-3 pathway by specific antibodies has been reported to efficiently inhibit experimental tumor lymphangiogenesis and metastasis in animal experiments. Although lymphangiogenesis has been reported to play an important role in the occurrence of colon cancer and to be associated with prognosis, it remains unclear whether it is a valid therapeutic target molecule. Further study of the potential of targeting this process for anti-lymphatic therapies is worthwhile.

Increased lymphangiogenesis in lung metastases from colorectal cancer is associated with early lymph node recurrence and decreased overall survival.

Pulmonary metastasectomy (PM) is an accepted treatment modality in colorectal cancer (CRC) patients with pulmonary tumor spread. Positive intrathoracic lymph nodes at the time of PM are associated with a poor prognosis and 5-year survival rates of <20%. Increased lymphangiogenesis in pulmonary metastases might represent an initial step for a subsequent lymphangiogenic spreading. We aimed to evaluate the presence of lymphangiogenesis in clinically lymph node negative patients undergoing PM and its impact on outcome parameters. 71 patients who underwent PM for CRC metastases were included in this dual-center study. Tissue specimens of pulmonary metastases and available corresponding primary tumors were assessed by immunohistochemistry for lymphatic microvessel density (LMVD) and lymphovascular invasion (LVI). Results were correlated with clinical outcome parameters. LMVD was 13.9±8.1 and 13.3±8.5 microvessels/field (mean±SD) in metastases and corresponding primary CRC; LVI was evident in 46.5 and 58.6% of metastases and corresponding primary CRC, respectively. Samples with high LMVD had a higher likelihood of LVI. LVI was associated with early tumor recurrence in intrathoracic lymph nodes and a decreased overall survival (p<0.001 and p=0.029). Herein, we present first evidence in a well-defined patient collective that increased lymphangiogenesis is already present in a subtype of pulmonary metastases of patients staged as N0 at the time of PM. This lymphangiogenic phenotype has a strong impact on patients' prognosis. Our findings may have impact on the post-surgical therapeutic management of CRC patients with pulmonary spreading.

SIRT1 expression is associated with lymphangiogenesis, lymphovascular invasion and prognosis in pN0 esophageal squamous cell carcinoma.

BackgroundSirtuin1 (SIRT1) is an NAD+-dependent type III histone deacetylase (HDAC). This research investigated the prevalence of SIRT1 protein expression and its prognostic influence with the aim of validating its potential role in lymphangiogenesis and lymphovascular invasion (LVI) in pN0 esophageal squamous cell carcinoma (ESCC).MethodsA total of 206 patients were enrolled in this retrospective study. SIRT1 and VEGF-C protein expression was detected by immunohistochemical staining. Peritumoral lymphatic microvessel density (LVD) and LVI were evaluated by immunostaining for D2-40. Statistical analysis was then preformed to investigate the relevance of SIRT1 expression and various clinicopathologic features and to examine the effect of SIRT1 on tumor-induced lymphangiogenesis, LVI and prognosis.ResultsSIRT1 positive expression was identified in 95 cases in the nucleus and was significantly correlated with T status (P < 0.001), disease stage (P = 0.001), VEGF-C positive expression (P = 0.015), high LVD (P = 0.013) and positive LVI (P = 0.015). Patients with SIRT1 positive expression, high LVD and positive LVI had a significantly unfavorable 5-year disease free survival (P < 0.001, P = 0.030, and P < 0.001, respectively) and overall survival (P < 0.001, P = 0.017, and P < 0.001, respectively). However, based on multivariate Cox regression analysis, only SIRT1 positive expression and positive LVI were significant independent prognosticators of poor disease-free survival (P = 0.029 and 0.018, respectively) and overall survival (P = 0.045 and 0.031, respectively).ConclusionsSIRT1 positive expression was significantly associated with tumor progression, lymphangiogenesis, LVI and poor survival in pN0 ESCC patients. Our research shows a utilization of SIRT1 in prognosing poor survival and providing possible target for ESCC patients through inhibiting its lymphangiogenesis activity.

Lymphatic microvessel density and vascular endothelial growth factor-C and -D as prognostic factors in breast cancer: a systematic review and meta-analysis of the literature

The use of lymphatic microvessel density (LVD) and pro-lymphangiogenic mediators as prognostic factors for survival in breast cancer remains controversial. We searched the electronic databases PubMed and EMBASE without language restrictions for relevant literature to aggregate the survival results. To be eligible, every study had to include the assessment of the LVD or the expression of vascular endothelial growth factor (VEGF)-C or -D in patients with breast cancer and provide a survival comparison, including disease-free survival (DFS) or overall survival (OS), according to the LVD, VEGF-C or VEGF-D status. Across all studies, 56.64 % of patients were considered to have a VEGF-C-positive tumor, and 65.54 % of patients had VEGF-D-positive tumors. High LVD had an unfavorable impact on DFS, with a pooled hazard ratio (HR) of 2.222 (95 % CI 1.579-3.126) and an OS with a HR of 2.493 (95 % CI 1.183-5.25). According to the different lymphatic makers, the subgroup HR in the D2-40 studies was 2.431 (95 % CI 1.622-3.644) for DFS and 4.085 (95 % CI 1.896-8.799) for OS. VEGF-C overexpression, as assessed by immunochemistry, was a prognostic factor for decreased DFS (HR 2.164; 95 % CI 1.256-3.729) and for decreased OS (HR 2.613; 95 % CI 1.637-4.170). VEGF-D overexpression was a significant although weak prognostic factor for DFS only when assessed by immunochemistry, with a HR of 2.108 (95 % CI 1.014-4.384). Our meta-analysis demonstrated that LVD, VEGF-C and VEGF-D could predict poor prognosis in patients with breast cancer. However, standardization of the assessment of LVD and for the expression of lymphangiogenesis factors is needed.

Peritumoral lymphatic microvessel density is related to poor prognosis in lung adenocarcinoma: A retrospective study of 65 cases

Although recent investigations have identified that lymphangiogenesis is associated with regional lymph node metastasis and tumor prognosis in non-small cell lung cancer (NSCLC), peritumoral lymphatic microvessel density (LMVD) and its prognostic significance in lung adenocarcinoma remain unknown. In the present study, we assessed peritumoral LMVD in lung adenocarcinoma and investigated its correlation with patient prognosis. Using immunohistochemistry (SP method), the D2-40-positive peritumoral LMVD count in lung adenocarcinoma was found to be 11.56±10.73, which was higher than intratumoral LMVD (P<0.001), and was found to be associated with lymphatic metastasis (P=0.003) and pTNM staging (P=0.046). Furthermore, a significant difference in the patient overall survival time was demonstrated between tumors with a high peritumoral LMVD and those with a low peritumoral LMVD (P=0.005). Finally, using multivariate analysis, it was determined that peritumoral LMVD, lymphatic metastasis and pTNM staging were independent prognostic factors. In conclusion, the results suggest that D2-40-positive peritumoral LMVD may predict the prognosis of lung adenocarcinoma.

Lymphangiogenesis as a Prognostic Marker in Breast Cancer Using D2-40 as Lymphatic Endothelial Marker—A Preliminary Study

We studied tumour lymphangiogenesis and lymphatic invasion using D2-40 endothelial marker in 35 breast cancer patients treated by primary surgery and correlated it with various clinico-pathological prognostic parameters. Lymphangiogenesis was quantified using lymphatic micro vessel density (LMVD) by counting the immunostained lymphatic microvessels at 200X. The mean age was 45.97±12.09 years (range 30-80 years). LMVD ranged from 5/hpf to 56/hpf with a mean score of 13.4±10.8 and median of 9. The median value of 9 was taken to classify patients into a low or high LMVD. LMVD correlated significantly with tumour size (p=0.003), histological grade (p=0.046), lymph node status (p=0.030). There was no significant correlation of LMVD with stage, estrogen receptor, progesterone receptor or HER2/neu immunoreactivity. Lymphovascular invasion on D2-40 staining [LVI-D40] was found in 13 (37.1%) cases compared to 6 cases (17.1%) on H & E staining showing a poor agreement (k=0.244). LVI correlated significantly with lymph node status (p=0.011). There was a strong association between tumour size (p=0.142), histological grade (p=0.066) though the correlation was not statistically significant. No correlation was found with stage, estrogen receptor, progesterone receptor or HER2/neu immunoreactivity. The mean LMVD in LVI positive patients was higher (22.85±13.29) as compared to LVI negative patients (7.95±2.05) and this was statistically significant (p=0.001). Increased D2-40 detected LMVD and LVI correlated with poor prognostic parameters.

Lymphatic microvessel density as a prognostic factor in non-small cell lung carcinoma: a meta-analysis of the literature

The role of lymphatic microvessel density (LVD) as a prognostic factor for survival of patients with non-small cell lung carcinoma (NSCLC) remains controversial. To evaluate this potential role, we performed a systematic review of the electronic databases PubMed and EMBASE for relevant literature to review and compile available survival results. To be eligible, a study had to assess LVD in patients with NSCLC and to compare survival based on LVD stratification. Among 12 eligible trials, all dealt with NSCLC, and 10 trials provided results for the meta-analysis of survival data (evaluable trials). In terms of survival, high LVD was reported to be an unfavorable prognostic factor for overall survival in 8 studies, whereas it was not in 4 studies. The overall survival hazard ratio for the 10 evaluable studies (1,426 patients) was calculated to be 1.41 (95% CI: 1.14-1.75) using a random effects model, indicating a poorer survival for NSCLC patients with high LVD. The hazard ratio was 1.52 (95% CI: 1.10-2.11) in 5 NSCLC studies where LVD was assessed based on D2-40 and 1.31 (95% CI: 1.08-1.60) in 4 studies where LVD was measured based on vascular endothelial growth factor receptor-3. This study supports the hypothesis that the lymphatic microvessel count or LVD, which reflects levels of lymphangiogenesis, is a poor prognostic factor for patient survival in surgically treated NSCLC. However, the present findings may overestimate the prognostic capacity of LVD because of publication and report bias. In addition, the standardization of lymphangiogenesis assessment by the lymphatic microvessel count is necessary.

Lymphangiogenesis in COPD: Another link in the pathogenesis of the disease

New lymphatic vessels are associated with tissue injury and repair. Recent studies have shown increased lymphatic follicles formation in the lungs of COPD patients. We hypothesized that lymphatic vascular remodeling could be part of COPD pathogenesis. To investigate the lymphangiogenetic process in COPD we measured the lymphatic microvessel density (LMVD), the lymphatic invasion (L.I), and their correlation with clinical and laboratory parameters. Lung tissue from 20 COPD patients and 20 non-COPD smokers was immunohistochemically stained for D2-40 (lymphatic endothelial cell marker), and LYVE-1 (lymphatic endothelial hyaluronan receptor 1). Both groups had similar age and smoking history. D2-40 and LYVE-1 were expressed in all specimens. Lymphatic invasion was presented only in COPD specimens. Lymphatic microvessel density (LMVD) as revealed by D2-40 and LYVE-1 markers was statistically significantly higher in COPD patients when compared with non-COPD smokers. Both markers (D2-40, LYVE-1) were correlated with FEV1 (% pred) (R(2)=0.415, R(2)=0.605, respectively). We report for the first time high lymphatic microvessel density and lymphatic invasion in COPD patients, related to the degree of airway obstruction. Our findings could provide novel insights in the pathogenesis of the disease.

Prognostic value of lymphangiogenesis in supraglottic laryngeal carcinoma

Metastasis to regional lymph nodes via lymphatic microvessels plays a key role in cancer progression, and is an important prognostic factor in many cancers. Recent evidence suggests that tumour lymphangiogenesis promotes lymphatic metastasis. To investigate whether tumour lymphatic microvessel density correlates with clinicopathological factors and serves as a prognostic indicator of supraglottic laryngeal carcinoma progression. The lymphatics of 84 supraglottic laryngeal carcinoma cases were investigated by immunohistochemical staining for podoplanin (also termed D2-40). The relationships between (intra- and peritumoural) lymphatic microvessel density, clinicopathological parameters and clinical prognosis were analysed. There was a significant relationship between high intratumoural lymphatic microvessel density and aggressive tumour node stage (p < 0.0001), distant metastasis (p = 0.037) and poor prognosis (p = 0.011), and between high peritumoural lymphatic microvessel density and node stage (p = 0.004) and poor prognosis (p = 0.029). Patients with high lymphatic microvessel density also had significantly worse disease-free survival (p = 0.003) and overall survival (p = 0.005). Intratumoural lymphatic microvessel density was found to be an independent prognostic factor for overall survival (p = 0.008) and disease-free survival (p = 0.005) (multivariate analysis). Lymphatic microvessel density (detected by podoplanin immunohistochemistry), especially intratumoural density, may be an independent predictor of lymphatic tumour spread and survival in supraglottic laryngeal carcinoma patients, and may be useful to guide decisions regarding additional surgery.

Abstract LB-289: Angiogenesis and lymphangiogenesis in mucoepidermoid carcinoma of the minor salivary glands

Abstract The purpose of this study was to investigate angiogenesis and lymphangiogenesis in mucoepidermoid carcinoma (MEC) of the minor salivary glands as potential predictors of tumor behavior. Intratumoral and peritumoral neoformed (NMVD) and lymphatic microvessel density (LMVD) were evaluated by CD105 and D2–40 immunohistochemistry, respectively. D2–40, VEGF-A, and VEGF-C immunoexpression were also investigated in tumor parenchyma. Twenty-six paraffin-embedded MEC samples were included in the study. Palate (50%) and buccal mucosa (19.2%) were the main affected sites. Sixteen (61.5%) tumors were low-grade, 6 (23.1%) intermediate-grade, and 4 (15.4%) high-grade. Twenty (77%) cases were clinically T1-T2, and 6 (23%) were T3-T4. Nodal metastasis occurred in 3 (11.5%) cases and distant metastasis in 1 (3.8%). Three (11.5%) cases recurred, and 3 (11.5%) patients died from disease. NMVD mean was 19.58±13.91 (400x field) for intratumoral evaluation and 10.90±7.79 (400x field) for peritumoral evaluation. LMVD mean was 5.66±3.43 (400x field) for peritumoral evaluation and 0.54±3.43 (400x field) for intratumoral evaluation. Just 6 (23.1%) cases reveal intratumoral lymphatic vessels. Twenty-two (84,6%) cases showed D2–40 in neoplastic cells, mainly in epidermoid type. Fifteen (57.7%) cases were positive for VEGFA, and 11 (42.3%) were positive for VEGF-C. A positive association was observed between VEGF-A positivity and intratumoral NMVD (p=0.05). Cases with nodal metastasis, altogether, were related to low intratumoral NMVD, high peritumoral LMVD, D2–40 expression and absence of VEGF-A. Distant metastasis was associated with low intratumoral NMVD, high peritumoral LMVD, D2–40 positivity and absence of VEGF-A and VEGF-C expression. Recurrent cases presented low intratumoral NMVD, and absence of VEGF-A and VEGF-C expression. Low intratumoral NMVD, and D2–40 positive cells were found in all tumors of patients that died of the disease. In conclusion, although NMVD, LMVD, D2–40, VEGF-A, and VEGF-C could not predict the prognosis of MEC, intratumoral NMVD, peritumoral LMVD, D2–40 and VEGF-A expression seems play a role in the occurrence of a worse behavior in this disease. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-289. doi:10.1158/1538-7445.AM2011-LB-289

Cytoplasmic HuR expression correlates with angiogenesis, lymphangiogenesis, and poor outcome in lung cancer

HuR is a ubiquitously expressed RNA-binding protein that stabilizes the mRNAs of certain genes and regulates the translation to proteins. Elevated cytoplasmic expression of HuR has been suggested to be associated with reduced survival in a wide variety of human carcinomas. However, the clinical significance of HuR expression in lung cancer remains unknown. In this study, we examined HuR expression in 132 patients with non-small cell lung carcinoma (NSCLC) by means of immunohistochemistry and correlated clinicopathologic data, lymphatic microvessel density (LVD), or microvessel density (MVD) with HuR immunostaining. HuR was expressed in 80.3% (106/132) of cases and was predominantly localized in the nucleus. Cytoplasmic HuR expression occurred in 40.9% (54/132) of NSCLC specimens and was associated with high MVD and LVD. In univariate analysis, cytoplasmic HuR, but not nuclear HuR expression was found to significantly influence the relapse-free survival and overall survival. In addition, cytoplasmic expression of HuR was identified as an independent prognostic factor for survival in multivariate analysis. Our data provide evidence for a clinically prognostic role of HuR in NSCLC and demonstrate an association between HuR expression, and angiogenesis and lymphangiogenesis.

Association between intratumoral lymphatic microvessel density (LMVD) and clinicopathologic features in endometrial cancer: a retrospective cohort study

BackgroundLymph node metastasis in endometrial cancer significantly decreases survival rate. Few data on the influence of intratumoral lymphatic microvessel density (LMVD) on survival in endometrial cancer are available. Our aim was to assess the intratumoral LMVD of endometrial carcinomas and to investigate its association with classical pathological factors, lymph node metastasis and survival.MethodsFifty-seven patients with endometrial carcinoma diagnosed between 2000 and 2008 underwent complete surgical staging and evaluation of intratumoral LMVD and other histologic variables. Lymphatic microvessels were identified by immunohistochemical staining using monoclonal antibody against human podoplanin (clone D2-40) and evaluated by counting the number of immunostained lymphatic vessels in 10 hot spot areas at 400× magnification. The LMVD was expressed by the mean number of vessels in these 10 hot spot microscopic fields. We next investigated the association of LMVD with the clinicopathologic findings and prognosis.ResultsThe mean number of lymphatic vessels counted in all cases ranged between 0 and 4.7. The median value of mean LMVD was 0.5, and defined the cut-off for low and high LMVD. We identified low intratumoral LMVD in 27 (47.4%) patients and high LMVD in 30 (52.6%) patients. High intratumoral LMVD was associated with lesser miometrial and adnaexal infiltration, lesser cervical and peritoneal involvement, and fewer fatal cases. Although there was lower lymph node involvement among cases with high LMVD, the difference did not reach significance. No association was seen between LMVD and FIGO staging, histological type, or vascular invasion. On the other hand, low intratumoral LMVD was associated with poor outcome. Seventy-five percent of deaths occurred in patients with low intratumoral LMVD.ConclusionOur results show association of high intratumoral LMVD with features related to more localized disease and better outcome. We discuss the role of lymphangiogenesis as an early event in the endometrial carcinogenesis.

Tumor-associated macrophages infiltration is associated with peritumoral lymphangiogenesis and poor prognosis in lung adenocarcinoma

Tumor-associated macrophages (TAMs) have been implicated in promoting tumor progression. Nowadays, adenocarcinoma has surpassed squamous cell carcinoma as the most frequent type of lung cancer, but in lung adenocarcinoma, the correlation of TAMs with lymphangiogenesis patients remains unclear. The aim of this study was to examine the relationship between TAMs and lymphangiogenesis and the lung adenocarcinoma patients' prognosis. Tumor specimens from 65 patients with lung adenocarcinoma were determined for TAMs count and lymphatic microvessel density (LMVD) by immunohistochemistry. A positive correlation existed between TAMs count and D2-40-positive peritumoral LMVD (r=0.069, P<0.001). TAMs infiltration was significantly associated with P-TNM staging (P=0.042) and lymph node metastasis (P=0.037), and peritumoral LMVD was correlated with lymph node metastasis (P=0.003). A significant difference in overall survival was detected not only between tumors with a high TAMs count and a low TAMs count (P=0.009) but also between tumors with a high peritumoral LMVD and a low peritumoral LMVD (P=0.005). Both TAMs count and peritumoral LMVD were independent prognostic factors for overall survival. Our results indicate that TAMs infiltration correlates with tumor lymphangiogenesis and poor survival in lung adenocarcinoma.

Prognostic significance of VEGF-C expression in correlation with COX-2, lymphatic microvessel density, and clinicopathologic characteristics in human non-small cell lung cancer

Lung cancer is one of the most lethal cancers in China because of high incidence and high mortality. Cyclooxygenase-2 (COX-2) and vessel endothelial growth factor C (VEGF-C) were found to play an important role in lymphangiogenesis of malignant tumors. In this study, we investigated whether lymphatic microvessel density (LMVD) is related to the prognosis in non-small cell lung cancer (NSCLC) patients, and the expressions of COX-2 and VEGF-C so as to determine the possible role of COX-2 and VEGF-C in NSCLC lymphangiogenesis. Sixty-five formalin-fixed paraffin embedded tissue samples of NSCLC were evaluated for COX-2 and VEGF-C by immunohistochemical staining. To assess tumor lymphangiogenesis, LMVD was determined by immunohistochemical staining of VEGFR-3 polyclonal antibody. The relationship among COX-2 and VEGF-C expression, LMVD, and clinicopathologic parameters was analyzed. Among the 65 samples, high LMVD was significantly associated with lymph node metastasis and poor survival. Multivariate survival analysis showed that LMVD value and lymph node metastasis were independent prognostic factors. The expression level of COX-2 and VEGF-C was significantly higher than those of the adjacent tissues. COX-2 and VEGF-C expressions in NSCLC significantly correlated with lymph node metastasis, but not with patient gender, age, tumor size, or tumor, nodes, metastasis classification stage. The mean LMVD value of COX-2- or VEGF-C-positive tumors was higher than that of COX-2- or VEGF-C-negative tumors. A significant correlation was found between the expressions of COX-2 and VEGF-C. This study suggests that LMVD may be one of the important prognostic factors for NSCLC patients. VEGF-C might play an important role in the COX-2 lymphangiogenic pathway. COX-2 and VEGF-C may play an important role in tumor progression by stimulating lymphangiogenesis. The inhibition of lymphangiogenesis, COX-2, or VEGF-C activity may have an important therapeutic benefit in the control of NSCLC.

Tumor-Associated Lymphangiogenesis Correlates with Prognosis after Resection of Human Hepatocellular Carcinoma

Experimental results from animal models as well as studies of human cancers indicate a critical role for tumor-associated lymphangiogenesis in tumor progression. However, its significance in hepatocellular carcinoma (HCC) is not well established. We analyzed tissue specimens from healthy liver (n = 36), cirrhotic liver (n = 24), and HCC (n = 60) by immunohistochemistry, using antibody D2-40 specific for lymphendothelia. We subsequently quantified lymphatic microvessel density (LVD). The LVD was correlated with clinicopathological characteristics of the tumors as well as survival and disease-free survival of the patients. In contrast to healthy as well as cirrhotic liver, lymphangiogenesis was induced in HCC. Lymphatic vessels were detected in the intratumoral septa as well as within the bulk of tumor cells. Tumors with high LVD (24 of 60) had developed significantly more frequently in cirrhotic livers (P = 0.001) and were more frequently restricted to one liver lobe (P = 0.04). Univariate analysis revealed high LVD as a marker for reduced survival and disease-free survival disadvantage (median >60 vs. 21 months, P = 0.018, and 19 vs. 8 months, P = 0.047, respectively). In multivariate analysis, LVD showed a trend toward association with reduced survival (P = 0.059) and represented an independent prognostic factor for disease-free survival (P = 0.017). Tumor-associated lymphangiogenesis is involved in neovascularization of hepatocellular carcinoma. Quantitative analysis of LVD demonstrated a significant influence of lymphangiogenesis on survival and established LVD as an independent predictor of disease-free survival. Quantification of LVD may be helpful in identifying patients with a high risk of tumor recurrence.

Tumor-Associated Lymphangiogenesis Correlates with Lymph Node Metastases and Prognosis in Hilar Cholangiocarcinoma

Tumor-associated lymphangiogenesis has been shown to promote nodal spread and is of prognostic significance in some tumor entities. Currently, nothing is known about the impact of lymphangiogenesis on progression and prognosis in hilar cholangiocarcinoma. We analyzed tissue specimens of normal liver and hilar cholangiocarcinoma (n = 60) by immunohistochemistry using the lymphendothelial-specific antibody D2-40 and subsequently quantified lymphatic microvessel density (LVD). The LVD was correlated with clinicopathological characteristics and recurrence pattern of the tumors as well as patients' survival. In contrast to the low abundance of lymphatic vessels in nontransformed liver tissue, we found an induction of lymphangiogenesis in hilar cholangiocarcinoma. Tumors with a high LVD (34 out of 60) had a significant higher incidence of lymph node involvement (p < 0.001), perivascular (p = 0.017), and perineural (p = 0.033) lymphangiosis and local recurrence (p < 0.001). Furthermore, a high LVD was identified to be a significant overall (three-year: 24.4% versus 90.5%; five-year: 7.0% versus 76.4%; p < 0.001) and disease-free (three-year: 8.3% versus 76.6%; five-year: 5.9% versus 61.4%; p < 0.001) survival disadvantage, with LVD representing an independent prognostic factor for survival (p < 0.001) in the multivariate analysis. Lymphangiogenesis is associated with increased frequency of tumor cells in lymphatics and lymph nodes in hilar cholangiocarcinoma. The prognostic importance of tumor-associated lymphangiogenesis was reflected by LVD serving as an independent prognostic factor. In addition, lymphangiogenesis may represent a potential target in the development of new therapeutic approaches in hilar cholangiocarcinoma.

Lymphatic Microvessel Density is an Independent Prognostic Factor in Colorectal Cancer

Although lymph node metastasis via lymphatic vessels often is related with an adverse outcome, it is not well known whether lymphatic spread to lymph node needs the development of the new lymphatic formation. In addition, the correlation between lymphangiogenesis and prognosis has not been well documented. This study was designed to assess the prognostic value of lymphangiogenesis and lymphatic vessel invasion in colorectal cancer. We examined 106 colorectal cancer specimens by immunostaining for podoplanin, lymphatic endothelial specific marker. We evaluated lymphangiogenesis, as measured by lymphatic microvessel density, and lymphatic vessel invasion. We next investigated the association of these two parameters with the clinicopathologic findings and prognosis. A significant correlation was observed between high lymphatic microvessel density and positive lymphatic vessel invasion (P = 0.0003). Positive lymphatic vessel invasion was significantly associated with the presence of lymph node metastasis (P = 0.0071). The survival curves demonstrated that both high lymphatic microvessel density and positive lymphatic vessel invasion were correlated with an adverse outcome (P = 0.0004 and P = 0.009, respectively). In a univariate analysis, high lymphatic microvessel density and positive lymphatic vessel invasion were negatively associated with the overall survival (P = 0.0011 and P = 0.0118, respectively). Furthermore, high lymphatic microvessel density, but not lymphatic vessel invasion, correlated with a poor outcome in a multivariate analysis (P = 0.0114). Our data suggested that lymphatic vessel invasion was related with lymph node metastasis and that both lymphatic microvessel density and lymphatic vessel invasion were related with an adverse outcome in colorectal cancer. Furthermore, lymphatic microvessel density may be a useful prognostic factor in colorectal cancer.

Lymphatic microvessel density using D2-40 is associated with nodal metastasis in non-small cell lung cancer

The monoclonal antibody D2-40 is a new selective marker for lymphatic endothelium. The lymphatic microvessel density (LMVD) using D2-40 has not yet been evaluated in non-small cell lung cancer (NSCLC). The aim of this study was to evaluate LMVD using D2-40 in NSCLC. We investigated LMVD in 77 patients with NSCLC who underwent curative tumor resection. We also determined the relation between LMVD and clinicopathologic factors, VEGF-C and Ang-2 and microvessel density (MVD) using factor VIII-related antigen. The median number of D2-40-positive vessels in the highest LMVD was 25 (range, 5-71). LMVD was significantly associated with tumor status, lymph node metastasis, stage, lymphatic invasion, VEGF-C protein and MVD (p=0.0149 for tumor status; p<0.0001 for nodal status; p<0.0001 for stage; p=0.0153 for lymphatic invasion; p=0.0030 for VEGF-C, and p=0.0029 for MVD). Furthermore, LMVD using D2-40 expression was shown to be an independent predictor of lymph node metastasis by multivariate analysis (p=0.0070). These data indicate that a high LMVD by D2-40 may be an indicator of lymph node metastasis in NSCLC.

Selective immunohistochemical staining shows significant prognostic influence of lymphatic and blood vessels in patients with malignant melanoma

Few data on the influence of lymphatic microvessel density (LMVD) on survival in patients with melanoma are available. The aim of this study was to assess LMVD and blood microvessel density (MVD) in tissue samples from 120 patients with melanoma. LMVD was stained with an antibody staining for podoplanin, and blood MVD was assessed by CD31 (PECAM-1)-immunostaining. Survival was determined using univariate and multivariate analysis. A significant association between a high CD31 MVD (but not LMVD) and the presence of lymph node metastases (P=0.007) was observed. Patients with a high LMVD had a significant shorter overall (OS) (P=0.0436) and disease-free survival (DFS) (P=0.0249) in univariate analysis. The survival analysis showed CD31 MVD was a strong prognostic factor for OS and DFS in both uni-and multivariate analyses. Our results demonstrate LMVD as a prognostic factor in malignant melanoma, although its prognostic relevance is much smaller compared with blood MVD.