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Objective To investigate the effect of exogenous vascular endothelial growth factor-C（VEGF-C）on the biological behavior of lung cancer cells and its mechanism. Methods We divided A549 cells into BK group, VC group, and SC group. A549 cells in the BK group were not transfected; the A549 cells in the VC group and SC group were added with 200 µL transfected liquid and 4 µL lipids into the EP tubes, and then A549 cells in the VC group and SC group were added with 5 µg NC mimics and MIR-21 mimics, respectively. The expression of TCAB1 protein in A549 cells was detected by Western blotting. The survival rate of A549 cells was detected by MTT. Transwell chamber was used to detect the inva⁃ sion of A549 cells. The apoptosis rate of A549 cells was detected by flow cytometry. Results The mRNA expression of VEGF-C in lung cancer tissues was lower in the SC group than in the BK group and the VC group（P<0. 05），which indi⁃ cated that the transfection was successful；the expression levels of VEGF-C mRNA in the BK group and the VC group were similar（P>0. 05）. There was no significant difference in the survival rate of A549 cells between the BK group and the VC group at 12，24，48，and 72 hours of intervention（P>0. 05）；the survival rate of A549 cells in the SC group was lower than those of the BK group and the VC group，and there was significant difference between the groups（P<0. 05）. There was no significant difference in the number of invasion cells between the BK group and the VC group（P>0. 05）. The num⁃ ber of invasion cells in the SC group was significantly lower than those of the BK group and the VC group；there were signif⁃ icant differences between the three groups（F=24. 383，P<0. 001）；there was no difference in the apoptosis rate between the BK group and the VC group（P>0. 05）. The apoptosis rate of A549 cells in the SC group was significantly higher than those of the BK group and the VC group；there were differences among the three groups（F=125. 326，P<0. 05）. No signif⁃ icant difference was found in the expression of TCAB1 protein in the lung cancer tissues between the BK group and VC group（2. 03 ± 0. 21 versus 1. 98 ± 0. 20）（P>0. 05），and the expression level of TCAB1 protein in A549 cells of the SC group（1. 35 ± 0. 12）was lower than those of the BK group，VC group（both P<0. 05）. Conclusion Reducing the level of exogenous VEGF-C can reduce the survival rate and invasion ability of A549 cells，and increase the apoptosis rate；its mechanism may be related to the inhibition of TCAB1 protein. [ABSTRACT FROM AUTHOR]

Lymphedema is a progressive and irreversible disease due to the lymphatic system disorder. Conservative and surgical therapies are either ineffective or impractical. Currently, mesenchymal stem cells (MSCs)-based therapies seem to be the most promising treatment for lymphedema. The MSCs promote lymphangiogenesis through the paracrine approach or by directly differentiating into lymphatic endothelial cells (LECs) under the induction of growth factors. Human dental pulp stem cells (hDPSCs) have been suggested to play important roles in tissue regeneration, making it an attractive candidate for the lymphedema treatment. In this study, to evaluate the potential role of hDPSCs in the clinical application for lymphedema treatment, we induced the hDPSCs with vascular endothelial growth factor-C (VEGF-C) and investigated the lymphangiogenic differentiation potential of hDPSCs in vitro. We found that under the VEGF-C induction, hDPSCs demonstrated upregulated LECs specific markers, promoted cell proliferation and migration, and increased tube formation, all of which contributed to their differentiation into LECs in vitro. [ABSTRACT FROM AUTHOR]

Middle-aged mice showed enhanced meningeal lymphatic function with MLVs expansion. Age-related changes in the function of MLVs are closely associated to the expression of VEGF-C in the cerebral cortex. Moreover, the cognitive function was enhanced in middle-aged mice compared to young and aged mice. These results indicate that the cerebral cortex may serve as a target for VEGF-C supplementation to ameliorate meningeal lymphatic dysfunction. MLVs, meningeal lymphatic vessels; VEGF-C, vascular endothelial growth factor-C. [Display omitted] • Middle-aged mice show improved meningeal lymphatic function and MLV expansion. • The middle-aged mice performed better in the Y maze test. • Age-related changes in meningeal lymphatic function are associated with VEGF-C expression. Meningeal lymphatic vessels (MLVs) have crucial roles in removing metabolic waste and toxic proteins from the brain and transporting them to the periphery. Aged mice show impaired meningeal lymphatic function. Nevertheless, as the disease progresses, and significant pathological changes manifest in the brain, treating the condition becomes increasingly challenging. Therefore, investigating the alterations in the structure and function of MLVs in the early stages of aging is critical for preventing age-related central nervous system degenerative diseases. We detected the structure and function of MLVs in young, middle-aged, and aged mice. Middle-aged mice, compared with young and aged mice, showed enhanced meningeal lymphatic function along with MLV expansion and performed better in the Y maze test. Moreover, age-related changes in meningeal lymphatic function were closely associated with vascular endothelial growth factor-C (VEGF-C) expression in the brain cortex. Our data suggested that the cerebral cortex may serve as a target for VEGF-C supplementation to ameliorate meningeal lymphatic dysfunction, thus providing a new strategy for preventing age-related central nervous system diseases. [ABSTRACT FROM AUTHOR]

Background: Secondary lymphoedema is a challenging pandemic. This condition may arise after oncologic resection of tumor-draining lymph nodes and/or radiation. Plastic-surgical procedures for lymphoedema comprise transplantation of vascularized lymph node flaps, which are, however, technically challenging and difficult to implement on a global level due to the scarcity of microsurgery facilities in some countries. To improve this situation, comparative research in valid animal models is needed. Methods: A total of 33 minipigs were subjected to lymphatic resection in the hind limbs. This large animal model was used in a first phase to compare different lymph node fragmentation methods and assess lymphatic regeneration after avascular transplantation. In a second phase, several stimulants were tested for their effect on lymphatic regeneration after fragment transplantation. In a third phase, animals additionally received irradiation of the groin. In this novel animal model, autologous avascular lymph node fragment transplantation was complemented by peripheral injections of vascular endothelial growth factor-C (VEGF-C). Finally, regeneration rates were quantified in relative numbers (percentage) in the irradiated tissue. Results: In the first phase, transversal lymph node fragmentation under preservation of the nodal capsule showed the best percentage of regeneration (62.5%). Peripheral intradermal administration of VEGF-C enhanced lymph node fragment regeneration (70.8%) better than injections of tetanus toxoid (41.6%) or Streptococcus suis (62.5%). Lymph node fragment regeneration also occurred in an irradiated porcine model of lymphadenectomy under VEGF-C administration (66.6%). Conclusions: The present findings provide a pre-clinical proof-of-concept for a possible simplification strategy for current operative procedures of autologous lymph node transplantation. Level of evidence : Not gradable [ABSTRACT FROM AUTHOR]

Objective To investigate whether vascular endothelial growth factor receptor-3 (VEGFR-3)/VEGF-C signaling suppresses activation of Kupffer cells (KCs) and attenuates hepatic ischemia-reperfusion injury (IRI) after liver transplantation. Methods Liver transplantation model was established in 12 pairs of Lewis-Lewis rats, 6 pairs for VEGF-C group and the other for control group. The donor livers of the VEGF-C group were perfused with VEGF-C injection via portal vein during cold preservation, while those of control group were perfused with UW solution. Another 6 rats served as sham operation group. In 24 h after transplantation, all rats were sacrificed, and serum levels of alanine transaminase (ALT), total bilirubin (TBIL) and inflammatory cytokines, as well as histological changes were detected and observed. KCs were isolated from the grafts. RT-PCR was used to evaluate the expression of VEGFR-3 and polarization-specific marker genes, EMSA was utilized to quantify the nuclear factor-κB (NF-κB) transcriptional activity, and Western blotting was employed to assess the expression of suppressor of cytokine signaling 1(SOCS1) and phosphorylated glycogen synthase kinase 3β (p-GSK3β). Results Compared with the controls, VEGF-C perfusion reduced serum ALT and TBIL levels, alleviated liver damage and suppressed the contents of proinflammatory cytokines (P < 0.05), but increased IL-10 level (P < 0.05). The VEGFR-3 mRNA level in KCs was increased after reperfusion in both VEGF-C group and control group (P < 0.05) when compared with the sham operation group (P < 0.05). In KCs, the mRNA levels of M1 specific marker genes and NF-κB activity were significantly inhibited in VEGF-C group (P < 0.05), while the mRNA levels of M2 specific marker genes and the expression of SOCS1 and p-GSK3β were enhanced when compared with control group (P < 0.05). Conclusion Exogenous VEGF-C protects liver graft from IRI by regulating the inflammatory responses and modifying polarization of KCs.

Objective To compare the effects of thoracic paravertebral nerve block-propofol intravenous balance general anesthesia (PPA) and sevoflurane inhalation balanced general anesthesia (SGA) on the serum concentrations of vascular endothelial growth factor C (VEGF-C) and transforming growth factor β1 (TGF-β1) in the patients undergoing radical resection for non-small cell lung cancer performed via video-assisted thoracoscopy. Methods Lung cancer patients undergoing radical resection were assigned to the PPA(n=23) or SGA(n=23) group. In the PPA group, thoracic paravertebral nerve block anesthetic was performed successfully with local injection of 0.5%ropivacaine(2.0 mg/kg) before induction of anesthesia, anesthesia was maintained with the target controlled infusion (TCI) of propofol (plasma concentration of 2.6~3.2 μg/mL)and intravenous remifentanil. In the SGA group, anesthesia was maintained with 1.0~1.5 (minimum alveolar concentration, MAC) sevoflurane and intravenous remifentanil. We observed opioid consumption intraoperative and 24 h postoperative, the pain scores at 2, 8, 24, 48, 72 h after surgery, as well as the serum concentrations of VEGF-C and TGF-β1 before surgery and 24 h after surgery. Results The amount of intraoperative remifentanil consumption in the PPA group was significantly lower than in the SGA group (P < 0.01), the amount of sufentanil consumption at 24 h postoperatively was significantly lower than in the SGA group (P < 0.01), and the VAS score at 2, 8 and 24 h postoperatively was significantly lower than in the SGA group (P < 0.01). At 24 h after surgery, the serum VEGF-C levels were lower in group PPA compared with the group SGA (629±203 vs 908±222 pg/mL, P=0.030), the serum TGF-β1 levels were lower in group PPA compared with the group SGA (8.37±2.84 vs 10.57±2.47 ng/mL, P=0.021). Conclusion Thoracic paravertebral nerve block-propofol intravenous balance general anesthesia can reduce opioids consumption for patients undergoing radical surgery for lung cancer, improve the postoperative analgesic effect, and reduce the concentration of serum tumor angiogenesis-related factors at 24 h after surgery. It is suggested that thoracic paravertebral nerve block-propofol intravenous balance general anesthesia is better than sevoflurane inhalation balanced general anesthesia for lung cancer patients undergoing radical resection.

AIM: To study the effects of Shuangdan Mingmu capsule on the expressions of vascular endothelial growth factor-a(VEGF-a), VEGF-b, VEGF-c in the retina of a diabetic rat model. METHODS: Forty male SD rats were divided into Group A(normal group), Group B(model group), Group C(Shuangdan Mingmu group)and Group D(positive control group)10 rats(20 eyes)in each group. A rat model of diabetic retinopathy was established by one-time tail vein injection with STZ(50mg/kg). After modeling for 1wk, the rats were given medicine by gavage. After gavage for 4wk rats were sacrificed, and the expressions of VEGF-a, VEGF-b, VEGF-c in the retina tissues were detected by immunohistochemical method. RESULTS: After gavage for 4wk the average gray values of VEGF-a, VEGF-b and VEGF-c protein in the retina of model group, Shuangdan Mingmu group and positive control group were lower than those of the normal group, and the average optical density were higher than those of the normal group. There was a significant difference between the model group and the normal group(PPPCONCLUSION: Shuangdan Mingmu capsule could significantly reduce the expressions of VEGF-a, VEGF-b,VEGF-c in the retina and had a certain protective effect on the retina of rats in the diabetic retinopathy model.

Background/Aims: Hyperglycemia has been shown to increase the incidence and metastasis in various types of cancers. However, the correlation between hyperglycemia and lymphatic metastasis in prostate cancer (PCa) remains unclear. Our previous study demonstrated that lysophosphatidic acid (LPA) enhances vascular endothelial growth factor-C (VEGF-C) expression, a lymphangiogenic factor, through activating it receptors LPA1/3 in prostate cancer (PCa) cells. Moreover, hyperglycemia up-regulates autotaxin (ATX) expression, a LPA-generating enzyme. Therefore, we propose that high glucose promotes VEGF-C expression through LPA signaling in PCa cells. Methods: Pharmacological inhibitors and siRNAs were utilized to investigate the molecular mechanism of high glucose-induced VEGF-C expression. Real-time PCR and Western blot were used to determine the mRNA and protein expressions, respectively. Cellular bioenergetics analysis was performed to determine the glycolysis levels. Results: We demonstrated that the expressions of VEGF-C, ATX, and calreticulin were increased upon high glucose treatments in PC-3 cells. Moreover, high glucose-induced VEGF-C expression was mediated through the LPA1/3, PLC, Akt, ROS and LEDGF-dependent pathways. Additionally, high glucose enhanced the aerobic glycolysis via LPA1/3. Conclusion: These results indicated that hyperglycemia leads to LPA synthesis, and subsequent promoting pathological consequence of PCa. These novel findings could potentially provide new strategies for PCa treatments.

The expression of pro-lymphangiogenic VEGF-C in primary tumors is associated with sentinel lymph node metastasis in most solid cancer types. However, the impact of VEGF-C on distant organ metastasis remains unclear. Perivascular tumor-associated macrophages (TAMs) play a crucial role in guiding hematogenous spread of cancer cells by establishing metastatic pathways within the tumor microenvironment. This process supports breast cancer cell intravasation and metastatic dissemination. We show here that VEGF-C-expressing TAMs reduce the dissemination of mammary cancer cells to the lungs while concurrently increasing lymph node metastasis. These TAMs express podoplanin and interact with normalized tumor blood vessels expressing VEGFR3. Moreover, clinical data suggest inverse association between VEGF-C-expressing TAMs and breast cancer malignancy. Thus, our study elucidates the paradoxical role of VEGF-C-expressing TAMs in redirecting cancer cells to preferentially disseminate to lymph nodes rather than to lungs, partially achieved by normalizing tumor blood vessels and promoting lymphangiogenesis. [Display omitted] • VEGF-C from TAMs increases metastases in the lymph nodes compared with the lungs • VEGF-C-expressing TAMs increase tumor vessel normalization and reduce hypoxia • Enforced expression of VEGF-C in TAMs enhances expression of podoplanin • Inverse association between VEGF-C-expressing TAMs and breast cancer progression In this study, Banerjee et al. show that VEGF-C expression from tumor-associated macrophages (TAMs) rewires the metastatic spreading of breast cancer cells by promoting vessel normalization and enhancing lymph angiogenesis. These changes lead to preferential dissemination of the cancer cells to the lymph nodes rather than to the lungs. [ABSTRACT FROM AUTHOR]

The lymphatic vasculature plays a key role in regulating tissue fluid homeostasis, lipid transport, and immune surveillance throughout the body. Although it has been appreciated that the heart relies on lymphatic vessels to maintain fluid balance and that such balance must be tightly maintained to allow for normal cardiac output, it has only recently come to light that the lymphatic vasculature may serve as a therapeutic target with which to promote optimal healing following myocardial ischemia and infarction. This article reviews the subject of cardiac lymphatic vessels and highlights studies that imply targeting of lymphatic vessel development or transport using vascular endothelial growth factor-C therapy may serve as a promising avenue for future clinical application in the context of ischemic injury.

Pure total flavonoids from Citrus (PTFC) effectively reduce the symptoms of non-alcoholic fatty liver disease (NAFLD). Our previous microarray analysis uncovered the alterations of important signaling pathways in the treatment of NAFLD with PTFC. However, the underlying core genes that might be targeted by PTFC, which play important roles in the progression of NALFD are yet to be identified. In this study, we predicted the vascular endothelial growth factor-C (VEGF-C) as potential key molecular target of PTFC against NAFLD via network pharmacology analysis. The network pharmacology approach presented here provided important clues for understanding the mechanisms of PTFC treatment in the development of NAFLD.

Vascular endothelial growth factor (VEGF)-C and its receptor, vascular endothelial growth factor receptor (VEGFR)-3, are responsible for lymphangiogenesis in both embryos and adults. In epilepsy, the expression of VEGF-C and VEGFR-3 was significantly upregulated in the human brains affected with temporal lobe epilepsy. Moreover, pharmacologic inhibition of VEGF receptors after acute seizures could suppress the generation of spontaneous recurrent seizures, suggesting a critical role of VEGF-related signaling in epilepsy. Therefore, in the present study, the spatiotemporal expression of VEGF-C and VEGFR-3 against pilocarpine-induced status epilepticus (SE) was investigated in C57BL/6N mice using immunohistochemistry. At 1 day after SE, hippocampal astrocytes and microglia were activated. Pyramidal neuronal death was observed at 4 days after SE. In the subpyramidal zone, VEGF-C expression gradually increased and peaked at 7 days after SE, while VEGFR-3 was significantly upregulated at 4 days after SE and began to decrease at 7 days after SE. Most VEGFC/VEGFR-3-expressing cells were pyramidal neurons, but VEGF-C was also observed in some astrocytes in sham-manipulated animals. However, at 4 days and 7 days after SE, both VEGFR-3 and VEGF-C immunoreactivities were observed mainly in astrocytes and in some microglia of the stratum radiatum and lacunosum-moleculare of the hippocampus, respectively. These data indicate that VEGF-C and VEGFR-3 can be upregulated in hippocampal astrocytes and microglia after pilocarpine-induced SE, providing basic information about VEGF-C and VEGFR-3 expression patterns following acute seizures. [ABSTRACT FROM AUTHOR]

Pure total flavonoids from Citrus (PTFC) effectively reduce the symptoms of non-alcoholic fatty liver disease (NAFLD). Our previous microarray analysis uncovered the alterations of important signaling pathways in the treatment of NAFLD with PTFC. However, the underlying core genes that might be targeted by PTFC, which play important roles in the progression of NALFD are yet to be identified. In this study, we predicted the vascular endothelial growth factor-C (VEGF-C) as potential key molecular target of PTFC against NAFLD via network pharmacology analysis. The network pharmacology approach presented here provided important clues for understanding the mechanisms of PTFC treatment in the development of NAFLD. [ABSTRACT FROM AUTHOR]

The association between homeobox (HOX)B9 expression and tumor malignancy was identified recently. It was reported that HOXB9 induced tumor angiogenesis, and associated with poor prognosis in patients with breast and colon cancer. On the other hand, regional lymph nodes are the most common site of tumor spread, and lymph node metastasis is a major prognostic factor in gastric cancer. It was hypothesized that HOXB9 promotes tumor lymphangiogenesis and induces tumor progression, invasion and metastasis in gastric cancer. The aim of the present study was to evaluate the correlation between HOXB9 expression, prognosis and clinicopathologic factors in patients with gastric cancer, and to assess the contribution of HOXB9 expression to tumor cell lymphangiogenesis in vitro. HOXB9 expression was evaluated by immunohistochemistry in resected tumor tissues from 58 patients with gastric cancer, and the association between prognosis and clinicopathologic factors was determined. HOXB9 gene was overexpressed in human gastric cancer TMK-1 cells and the effect of HOXB9 overexpression on the expression of vascular endothelial growth factor (VEGF)-C, VEGF-D and VEGF receptor (R)-3 was determined. It was demonstrated that the depth of tumor invasion, the number of node metastases, lymphatic invasion and vascular invasion were significantly associated with HOXB9 expression. Overall survival was decreased in patients with HOXB9 expression. The mRNA expression of VEGF-D but not of VEGF-C and VEGFR-3 was increased in HOXB9-overexpressing TMK-1 cells compared with control cells. In conclusion, HOXB9 expression was positively correlated with gastric cancer progression and lymphangiogenesis marker expression. HOXB9 may be associated with lymphogenic metastasis. [ABSTRACT FROM AUTHOR]

Objective To observe the expression changes of CD133 and vascular endothelial growth factor-C( VEGFC) in the oral squamous cell carcinoma( OSCC) and to analyze the clinical significance. Methods Totally 163 OSCC paraffin-embedded tissues were collected as the observation group, 37 cases of normal oral mucosa paraffin-embedded tissues were collected as the control group. Immunohistochemical method was used to detect the expression of CD133 and VEGF-C in both of the two groups, meanwhile, the lymphatic vessel density( LVD) of two groups were also measured, and then we analyzed the correlation between CD133 and VEGF-C through Spearman rank correlation analysis. In addition, the relationships of CD133 and VEGF-C with clinical pathological parameters in patients with OSCC were also analyzed. [ABSTRACT FROM AUTHOR]

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Background/purpose: Evidence shows that there is a relationship between hypoxia and inflammatory response in periodontitis. Hypoxia-inducible factor (HIF)-1α is a major regulator of energy homeostasis and cellular adaptation to low oxygen stress. Although experimental results demonstrate an association between HIF-1α and vascular endothelial growth factor (VEGF)-C in tumor angiogenesis, the role of HIF-1α and VEGF-C in the pathogenesis of periodontitis is still ambiguous. So far, limited attention has been given to the role of hypoxia and VEGF-C in periodontitis. The present study aimed to investigate the expression and distribution of HIF-1α and VEGF-C in gingival tissue samples from patients with different stages of chronic periodontitis and healthy individuals. Materials and methods: A total of 56 samples were involved in this study, including moderate chronic periodontitis (n = 20), advanced chronic periodontitis (n = 20), and healthy control tissues (n = 16). The gingival specimens were stained with hematoxylin and eosin for histopathology. The expression of HIF-1α and VEGF-C in gingival tissues was detected by immunohistochemical staining. Results: HIF-1α and VEGF-C were found in gingival tissues from patients with different stages of chronic periodontitis as well as healthy control tissues. HIF-1α protein was expressed mainly in the epithelial layer of gingival tissues, and VEGF-C protein was mostly located in the connective tissue papilla of gingival tissues. Compared with healthy controls, the expression of HIF-1α and VEGF-C in chronic periodontitis groups was significantly higher (P

Background/Aims: Hyperglycemia has been shown to increase the incidence and metastasis in various types of cancers. However, the correlation between hyperglycemia and lymphatic metastasis in prostate cancer (PCa) remains unclear. Our previous study demonstrated that lysophosphatidic acid (LPA) enhances vascular endothelial growth factor-C (VEGF-C) expression, a lymphangiogenic factor, through activating it receptors LPA1/3 in prostate cancer (PCa) cells. Moreover, hyperglycemia up-regulates autotaxin (ATX) expression, a LPA-generating enzyme. Therefore, we propose that high glucose promotes VEGF-C expression through LPA signaling in PCa cells. Methods: Pharmacological inhibitors and siRNAs were utilized to investigate the molecular mechanism of high glucose-induced VEGF-C expression. Real-time PCR and Western blot were used to determine the mRNA and protein expressions, respectively. Cellular bioenergetics analysis was performed to determine the glycolysis levels. Results: We demonstrated that the expressions of VEGF-C, ATX, and calreticulin were increased upon high glucose treatments in PC-3 cells. Moreover, high glucose-induced VEGF-C expression was mediated through the LPA1/3, PLC, Akt, ROS and LEDGF-dependent pathways. Additionally, high glucose enhanced the aerobic glycolysis via LPA1/3. Conclusion: These results indicated that hyperglycemia leads to LPA synthesis, and subsequent promoting pathological consequence of PCa. These novel findings could potentially provide new strategies for PCa treatments. [ABSTRACT FROM AUTHOR]

Background: Recent studies, mainly on animal models, have suggested that negatively charged glycosaminoglycans, macrophages, and lymph vessels in the skin interstitium may serve as extrarenal control of sodium balance and blood pressure. The aim of the study was to prove the hypothesis that skin interstitium has a role in the pathogenesis of hypertension in humans. Methods and Results: We have examined skin biopsies in 91 patients from the department of surgery who had elective surgery with abdominal skin incision: 43 were hypertensive, 14 had resistant hypertension, and 34 with normal blood pressure as control group (median patients' age in these groups estimated accordingly 64 vs. 64 vs. 61.5; p > 0.05). We have studied (1) the content of Na+, water, accumulation of macrophages (CD68), and density of lymphatic vessels (D2-40) and blood vessels (CD31) in the specimens of abdominal skin taken at the time of surgery and (2) plasma NT-proANP, vascular endothelial growth factor (VEGF)-C, and VEGF-D concentrations. The study groups differed in skin expression of CD68 (control vs. hypertension vs. resistant hypertension groups were accordingly: 3.33 vs. 4.00 vs. 8.33; p = 0.005) and in serum concentration of VEGF-C (5792 vs. 4348 vs. 3974 pg/mL; p = 0.026). Differences among groups in plasma NT-proANP levels were close to statistical significance (p = 0.056). Conclusions: Our results confirm that skin interstitium may be involved in the pathogenesis of essential hypertension in humans. Lower levels of VEGF-C in hypertensive groups suggest that impairment of lymphangiogenesis and protective function of the skin lymphatic system may play a role in the pathogenesis of hypertension. [ABSTRACT FROM AUTHOR]

Purpose: Papillary thyroid carcinoma (PTC) has a strong propensity to metastasize to regional lymph nodes which increases the risk of local-regional relapse and affects the course of the disease. Molecular pathogenesis of lymph node metastasis (LNM) is not yet fully understood. Survivin, a multifunctionale molecule involved in apoptosis, proliferation and angiogenesis, and vascular endothelial growth factor-C (VEGF-C) are suggested to be implicated in lymphatic metastases of human malignancies.Materials and Methods: Expression of survivin and VEGF-C was examined by immunohistochemistry and Western blot in 75 cases of PTCs in relation to their LNM status. Additionally, survivin and VEGF-C were immunohistochemically analyzed in 15 primary PTCs paired with their metastatic tissue in lymph nodes.Results: High expression of survivin and VEGF-C was found in 62.7% and 64.0% cases, respectively, with a positive correlation to each other (Spearman's correlation co-efficient = 0.878, P < 0.001). Expression levels of both proteins were significantly higher in patients with LNM than in those without LNM (P < 0.001). The rate of concomitant high expression of survivin and VEGF-C in patients with LNM involvement was 88.9% (P < 0.01). Metastatic tissue in lymph nodes expressed survivin and VEGF-C at the same high extent as their primary tumors.Conclusion: Concomitant high expression of survivin and VEGF-C is closely associated with LNM status of PTC patients, which suggests their cooperation in the metastatic process. Evaluation of survivin and VEGF-C expression could be clinically significant in predicting the metastatic potential of PTC and subsequent treatment and follow-up of these patients. [ABSTRACT FROM AUTHOR]

Aim: To evaluate the expression of two different lymphatic vascular density (LVD) markers (D2-40 and LYVE-1) and a lymphangiogenic cytokine (Vascular Endothelial Growth Factor-C, [VEGF-C]) in prostate carcinoma and to investigate their relationship with the lymph node status. Settings and Design: Archival material study of 92 non-consecutive radical prostatectomy specimens. Materials and Methods: The mean LVD was assessed immunohistochemically in 24 prostate carcinoma specimens from patients with clinically localized disease, who were found to have nodal metastasis (pN1), and was compared with 68 pN0 cases. Furthermore, the mean LVD, VEGF-C expression, and lymphatic invasion were examined in relation to lymph node involvement. Results: Peritumoral (but not intratumoral) mean LVD assessed by D2-40 was higher in pN1 tumors (P = 0.015). LYVE-1 expression was limited and not associated with lymph node status. The VEGF-C expression was higher in the N1 cases and also correlated with the increased mean LVD in both the peri- and intratumoral compartments. Lymphatic invasion was strongly associated with nodal metastasis and higher VEGF-C expression. Conclusions: Our results indicate that increased peritumoral (but not intratumoral) LVD in the tumor specimen is associated with lymph node metastasis. Increased expression of VEGF-C is associated with higher LVD (in both intratumoral and peritumoral compartments) and with positive lymph node status, indicating a possible dual role in both lymphangiogenesis and lymphatic vessel invasion.

Temozolomide (TMZ) is an alkylating agent used for the treatment of aggressive forms of brain tumor based on its antitumor actions. However, the exact effect on cancer and the underlying anticancer molecular mechanism of TMZ remain to be elucidated. In the present study, the effects of TMZ on the growth and motility of glioma C6 cells were investigated. MTT and Transwell assays were used to detect cellular growth and motility. The results showed that TMZ inhibited the proliferation, migration and invasion of the glioma C6 cells in vitro, western blot analysis determined that the phosphorylation of extracellular signal-regulated protein kinase (ERK)1/2 was decreased in the TMZ-treated cells, compared with the untreated control cells. The ERK1/2 specific inhibitor, U0126, augmented the inhibitory effects of TMZ on the proliferation, migration and invasion of the glioma C6 cells, and the mitogen-activated protein kinase kinase/ERK pathway activator, curcumin, attenuated the inhibitory effects of TMZ on the proliferation and motility of the glioma C6 cells. Additionally, the western blotting in the present study demonstrated that TMZ and U0126 decreased the expression of vascular endothelial growth factor-C (VEGF-C), and the expression level was restored by curcumin, suggesting that VEGF-C may be the downstream effector of ERK1/2. Furthermore, the overexpression of VEGF-C enhanced the growth, migration and invasion of the TMZ-treated cells. These results suggested that TMZ suppressed glioma C6 cell development, at least in part, and downregulated the expression of VEGF-C by inhibiting the ERK signaling pathway. The results of the present study provides the foundation for a combinational therapeutic strategy to improve the efficacy of TMZ. [ABSTRACT FROM AUTHOR]

Vascular endothelial cell growth factor (VEGF)-C promotes tumorigenesis by allowing lymph node metastasis and lymphangiogenesis, among other actions. RNA interference (RNAi) is a novel technique for suppressing target gene expression and may increase the effectiveness of cancer treatments. The present study assessed the influence of VEGF-C RNAi on the apoptosis and proliferation of mouse breast cancer cells in vitro and in vivo. A total of three pairs of small interfering RNA (siRNA) targeting mouse VEGF-C were designed and synthesized prior to transfection into 4T1 cells via a liposomal approach. Reverse transcription polymerase chain reaction, western blot analysis, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Hoechst 33258 staining and flow cytometry were performed in vitro to analyze VEGF-C expression, cleaved caspase-3 protein expression and 4T1 cell proliferation and apoptosis. Experiments were also conducted in vivo on BALB/c mice with breast cancer. Tumor weight and volume were measured and the number of apoptotic cells in tumor tissues was assessed by a TUNEL assay. Immunohistochemical assays and an enzyme-linked immunosorbent assay were used to measure the expression of VEGF-C in tumor tissues. The results demonstrated that the three pairs of siRNA, particularly siV2, significantly reduced VEGF-C mRNA and protein levels in 4T1 cells. siV2 was deemed to be the most efficient siRNA and therefore was selected to be used in subsequent experiments. Furthermore, in vitro studies indicated that VEGF-C RNAi significantly decreased cell growth, induced apoptosis and upregulated the expression of cleaved caspase-3 protein. Tumor weight and volume in breast cancer in vivo models was reduced by the intratumoral injection of siV2. Antitumor efficacy was associated with decreased VEGF-C expression and increased induction of apoptosis. The present study therefore indicated that VEGF-C RNAi inhibited mouse breast cancer growth in vitro and in vivo and that it may be a novel targeted therapy for breast cancer. [ABSTRACT FROM AUTHOR]

Background and objective Vascular endothelial growth factor-C (VEGF-C), a member of the VEGF family, has been proven to be a relatively special VEGF. When binding to its receptor VEGFR-3, it activates lymphangiogenesis, which likely induces lymph node metastasis in tumors. The aim of this study is to characterize the expression and prognostic value of VEGF-C and lymphangiogenesis in lung adenocarcinoma and squamous cell carcinoma. Methods Through immunohistochemistry, the lymph vessel endothelial cells undergoing lymphangiogenesis were stained with podoplanin to detect the expression of VEGF-C and lymphangiogenesis in 98 cases with stage IIIa (N2) lung adenocarcinoma and squamous cell carcinoma. Results The expression rate of the VEGF-C was positively correlated with the lymphatic microvessel density (LMVD; r=0.783, P

Background and objective It has been proven that some molecular markers play important roles in diagnosis, clinical prediction and prognosis monitoring for lung cancer. The aim of this study is to investigate the expressions and their significances of vascular endothelial growth factor-C (VEGF-C) mRNA and epidermal growth factor receptor (EGFR) mRNA in non-small cell lung cancer (NSCLC) tissues and lymph node tissues. Methods The expressions of VEGF-C mRNA and EGFR mRNA were detected in tumor tissues and lymph node tissues by reverse transcription polymerase chain reaction (RT-PCR) and real-time fluorescence quantitative PCR. The relationship and correlation between the expressions of VEGF-C mRNA, EGFR mRNA and clinicopathologic characteristics were analyzed. Results The expressions of VEGF-C mRNA and EGFR mRNA in tumor tissues and lymph node tissues were significantly higher than those in control group (P < 0.05). The expressions of VEGF-C mRNA and EGFR mRNA were much higher in patients with lymph node metastasis than those without lymph node metastasis (P < 0.05). The expressions of VEGF-C mRNA and EGFR mRNA in tumor tissues and lymph node tissues were significantly correlated (r=0.834, P < 0.001; r=0.817, P < 0.001). Conclusion The overexpressions of VEGF-C and EGFR genes are closely related to lymph node metastasis. VEGF-C mRNA and EGFR mRNA are valuable biomarkers for early diagnosis, clinical prediction and prognosis for lung cancer.

Lymphatic vessels drain excess tissue fluids to maintain the interstitial environment. Lymphatic capillaries develop during the progression of tissue fibrosis in various clinical and pathological situations, such as chronic kidney disease, peritoneal injury during peritoneal dialysis, tissue inflammation, and tumor progression. The role of fibrosis-related lymphangiogenesis appears to vary based on organ specificity and etiology. Signaling via vascular endothelial growth factor (VEGF)-C, VEGF-D, and VEGF receptor (VEGFR)-3 is a central molecular mechanism for lymphangiogenesis. Transforming growth factor-β (TGF-β) is a key player in tissue fibrosis. TGF-β induces peritoneal fibrosis in association with peritoneal dialysis, and also induces peritoneal neoangiogenesis through interaction with VEGF-A. On the other hand, TGF-β has a direct inhibitory effect on lymphatic endothelial cell growth. We proposed a possible mechanism of the TGF-β–VEGF-C pathway in which TGF-β promotes VEGF-C production in tubular epithelial cells, macrophages, and mesothelial cells, leading to lymphangiogenesis in renal and peritoneal fibrosis. Connective tissue growth factor (CTGF) is also involved in fibrosis-associated renal lymphangiogenesis through interaction with VEGF-C, in part by mediating TGF-β signaling. Further clarification of the mechanism might lead to the development of new therapeutic strategies to treat fibrotic diseases.

Background and objective Heparanase has been thought to be a good molecular marker of tumor, and the heparanase expression level was correlated closely with tumor metastasis. In this study, we investigate the effects of heparanase on angiogenesis and lymphangiogenesis of lung cancer and the relationship between heparanase expression and vascular endothelial growth factor (VEGF), vascular endothelial growth factor-C (VEGF-C). Methods Immunohistochemistry was used to detect the expression of heparanase, VEGF, VEGF-C protein and microvascular density (MVD), lymphatic vessel density (LVD) in 115 cases of non-small cell lung cancer (NSCLC) and 45 cases of adjacent normal tissue samples. Results Our results showed that heparanase expression was significantly increased in 91 (79.13%) of the 115 cases and correlated with lymph node metastasis (node positive rate 87.0%; node negative rate 36.8%; P=0.003). Heparanase positive expression cases have significantly higher concentration of microvascular density (MVD) and lymphatic vessel density (LVD) as compared with heparanase negative expression cases (P

The lymphatic vascular system is crucial for the regulation of tissue fluid homeostasis, lipid metabolism, and immune function. Cardiac injury quickly leads to myocardial edema, cardiac lymphatic dysfunction, which ultimately results in myocardial fluid imbalance and cardiac dysfunction. Therefore, lymphangiogenesis-targeted therapy may improve the recovery of myocardial function post cardiac ischemia as observed in myocardial infarction (MI). Indeed, a promising strategy for the clinical treatment of MI relies on vascular endothelial growth factor-C (VEGF-C)-targeted therapy, which promotes lymphangiogenesis. However, much effort is needed to identify the mechanisms of lymphatic transport in response to heart disease. This article reviews regulatory factors of lymphangiogenesis, and discusses the effects of lymphangiogenesis on cardiac function after cardiac injury and its regulatory mechanisms. The involvement of stem cells on lymphangiogenesis was also discussed as stem cells could differentiate into lymphatic endothelial cells (LECs) and stimulate phenotype of LECs. • The development of mature lymphatic network depends on the interaction of Prox-1, FOXC2, VEGFR-3, LYVE-1 and Podoplanin. • Lymphatics are immunomodulators, affect angiogenesis and modulate cardiac edema after myocardial infarction. • The number and morphology of lymphatic vessels were changed dynamically after myocardial infarction. • Stem cells differentiate into LECs and promote their proliferation, migration and formation of tubular structures. [ABSTRACT FROM AUTHOR]

Background We sought to comprehensively summarize available evidence for the use of VEGF-C protein to evaluate the clinicopathological and prognostic role of VEGF-C in colorectal cancer. Methods Electronic databases from inception to February 2016 were used to search without language restrictions for original articles. A meta-analysis was undertaken to assess the relationship between VEGF-C expression and overall survival (OS) in colorectal cancer. Results Twenty-seven studies were included in the final meta-analysis. We aggregated 13 trials ( n = 1.428 patients) that evaluated the correlation between OS and VEGF-C overexpression. Statistics were performed for OS (HR = 1.95; 95%CI = 1.31–2.92, P = 0.007). When the studies were stratified by the pathological variables, including T stage ( n = 383 patients; OR = 1.79; 95%CI = 1.14–2.81), lymph node metastasis ( n = 3212 patients; OR = 4.21; 95%CI = 3.49–5.08), M stage ( n = 1106 patients; OR = 4.46; 95%CI = 2.96–6.70), vascular invasion( n = 1471 patients; OR = 2.18; 95%CI = 1.65–2.88), lymph invasion ( n = 831 patients; OR = 3.95; 95%CI = 2.80–5.56), histo-differentiation ( n = 1695 patients; OR = 1.34; 95%CI = 1.00–1.79) and Duke's stage( n = 778 patients; OR = 4.90; 95%CI = 3.55–6.75), TNM stage ( n = 808 patients; OR = 1.73; 95%CI = 1.18–2.54) provided critical and comprehensive prognostic information. Conclusion Our results demonstrated that VEGF-C overexpression was associated with OS in colorectal cancer. [ABSTRACT FROM AUTHOR]

Introduction: The aim of this study is to determine whether regulation of the expression level of fms-like tyrosine kinase-4 (Flt-4) is related to osteoclast differentiation.Material and Methods: Osteoclast formation and differentiation of mouse bone marrow cells and RAW264.7 cells were performed. To induce osteoclast differentiation, RANKL (50 ng/ml) with or without vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor-D (VEGF-D) was added to mouse bone marrow cells and RAW264.7 cells. Then cells were examined under a microscope. TRAP-positive cells with 3 nuclei or more were considered as osteoclasts and counted. The Flt-4 gene was knocked down by transfection of siRNAs against Flt-4. Immunoblot analyses were performed.Results: The osteoclast formation assay indicated that VEGF-C resulted in 500 or 450 vs. 100 (p < 0.05) of osteoclasts in mouse bone marrow cells and RAW264.7 cells, respectively. Vascular endothelial growth factor-D resulted in about 600 or 630 vs. 100 (p < 0.05) of osteoclasts for both mouse bone marrow cells and RAW264.7 cells. The knock-down of Flt-4 expression abolished the induction by VEGF-C or VEGF-D, resulting in induction similar to that of the negative control PBS.Conclusions: Both VEGF-C and VEGF-D can induce osteoclast differentiation in the presence of the receptor activator of nuclear factor κB ligand. Down-regulation of expression level of Flt-4 protein abolishes osteoclast differentiation induced by VEGF-C or VEGF-D. [ABSTRACT FROM AUTHOR]

Vascular endothelial growth factor-C (VEGF-C) is a secreted growth factor involved in many oncogenic processes, and neuropilin-2 (NRP2) is essential for neuronal guidance as a well-acknowledged co-receptor of VEGF receptors. The overexpression of NRP2 has been reported in many types of cancers, but the significance of NRP2 in glioblastoma is not elucidated. To investigate the clinical significance of VEGF-C and NRP2 in glioblastoma, we detected their expression in 86 cases of glioblastoma with immunohistochemistry and categorized our cohort into high- and low-expression groups according to the immunohistochemistry score, which was the product of the score of staining intensity multiplied by the score reflecting the percentage of positive cells. The proportion of glioblastoma with high VEGF-C expression was 34.9% (30/86), and that with high NRP2 expression was 37.2% (32/86). The proportion of glioblastoma with high expression of both VEGF-C and NRP2 was 15.1% (13/86). Moreover, the proportion of cases with high VEGF-C and low NRP2 was 19.7% (17/86), while the proportion of cases with low VEGF-C and high NRP2 was 22.1% (19/86). The high expression of both VEGF-C and NRP2 was associated with poorer survival rate (P = 0.023), and can be identified as an independent prognostic factor in glioblastoma (P = 0.030). Moreover, cases with low NRP2 expression are easier for complete tumor resection (P = 0.038). In conclusion, the concurrent high expression of VEGF-C and NRP2 is predictive of the unfavorable prognosis in glioblastoma, indicating that the VEGF-C-NRP2 signaling pathway is a potential and promising drug target in glioblastoma therapy. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are a class of non-coding RNAs 18-25 nucleotides in length, which play important roles in the regulation of cancer progression through gene silencing. miRNA (miR)-101 has been suggested to be associated with hepatocellular carcinoma (HCC). However, the detailed role of miR-101 in HCC metastasis and the underlying mechanism remain largely unclear. The present study demonstrated that the expression of miR-101 was significantly reduced in HCC tissues compared with that in matched normal adjacent tissues. miR-101 was also found to be downregulated in four HCC cell lines compared with its expression in a normal liver cell line. Vascular endothelial growth factor (VEGF)-C was further identified as a direct target of miR-101, and the protein expression of VEGF-C was downregulated by miR-101 in HepG2 HCC cells. Furthermore, the overexpression of miR-101 and the knockdown of VEGF-C significantly inhibited HepG2 cell migration and invasion, while restoration of VEGF-C reversed the inhibitory effect of miR-101 overexpression on HepG2 cell migration and invasion. Finally, the expression of VEGF-C was notably increased in HCC tissues and cell lines. These findings suggest that miR-101 exerts a suppressive effect on HCC cell migration and invasion, at least in part through the direct inhibition of VEGF-C protein expression. Therefore, the miR-101/VEGF-C axis may serve as a potential therapeutic target for HCC metastasis. [ABSTRACT FROM AUTHOR]

We unveiled the association of Annexin A7 with vascular endothelial growth factor-C (VEGF-C) and the effect of upregulation of Annexin A7 in Hca-F and Hca-P cells on inhibiting hepatocarcinoma (HCC) lymph node metastasis (LNM) in vitro and in vivo. A total of 200 inbred 615 mice were randomly divided into four equal groups inoculated with Hca-F, Hca-P, F, and P cells, respectively. The primary tumor, popliteal, inguinal, and iliac lymph nodes were prepared for immunohistochemical (IHC) staining, real-time quantitative polymerase chain reaction (qRT-PCR) analysis, Western blot, and hematoxylin-eosin (H&E) staining. There was over 50 % increase both in the number of F and P cells migrated through the filter compared to their controls (F, Hca-F and P, Hca-P). However, no significant differences were noted in invasion ability between them (all P > 0.05). Tumor lymph vessels were significantly reduced in F and P tumors when compared with Hca-F and Hca-P tumors (all P < 0.05). Blood vessel density did not differ significantly between F and P tumors and Hca-F and Hca-P tumors. Enzyme-linked immunosorbent assay (ELISA) for VEGF-C showed that upregulating Annexin A7 decreased VEGF-C secretion in F and P cells ( P < 0.05). The IHC staining result showed that the level of serum Annexin A7 was found to be statistically higher in all experimental groups than that in the control group ( P < 0.05). The present results indicated that alterations in serum Annexin A7 expression may be of prognostic relevance in HCC lymphatic metastasis. [ABSTRACT FROM AUTHOR]

The objective of this study is to investigate relationship between the expressions of heparanase and vascular endothelial growth factor-C (VEGF-C) mRNA and tumorigenesis, progression in human lung cancer. The expressions of heparanase and VEGF-C mRNA in 65 cases of lung cancer (31 cases of squamous cell carcinoma, 25 adenocarcinoma, 3 large cell carcinoma, and 6 small cell carcinoma), adjacent tissues of cancer, and normal tissues were tested by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed by clinico-pathological characteristics and prognosis of lung cancer. The rate of expressions of heparanase and VEGF-C mRNA in tumor tissues (55.4, 61.5 %) was significantly higher than that in adjacent tissues of cancer (12.3, 15.4 %) and normal tissues (3.1, 4.6 %) ( P < 0.05). It was shown that heparanase and VEGF-C mRNA expressions did not correlate with the pathological type and grade of the tumor ( P > 0.05), but they correlated with the clinical stage and survival time of the patients ( P < 0.05). Overexpression of heparanase and VEGF-C mRNA in lung cancer tissues perhaps participates in regulation of tumorigenesis and progression. The expressions of heparanase and VEGF-C mRNA should be used as a useful marker of the biological behavior of lung cancer and as an independent prognosis factor for the patient's survival. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRs) have important roles in the pathogenesis of human malignancy. It has previously been suggested that deregulation of miR-101 is associated with the progression of intrahepatic cholangiocarcinoma (ICC); however, the exact role of miR-101 in the regulation of ICC metastasis remains largely unknown. The present study demonstrated that the expression levels of miR-101 were significantly decreased in ICC tissue, as compared with matched adjacent normal tissue. Furthermore, miR-101 was downregulated in the ICC-9810 human ICC cell line, as compared with in the normal human intrahepatic biliary epithelial cell (HIBEC) line. Vascular endothelial growth factor (VEGF)-C was identified as a target gene of miR-101 in ICC-9810 cells. The expression of VEGF-C was negatively regulated by miR-101 at the post-transcriptional level in ICC-9810 cells. Further investigation demonstrated that overexpression of miR-101 markedly suppressed the migration and invasion of ICC-9810 cells, and these effects were similar to those observed following VEGF-C knockdown. Conversely, restoration of VEGF-C reversed the inhibitory effects of miR-101 overexpression on ICC-9810 cell migration and invasion, thus suggesting that miR-101 may suppress ICC-9810 cell migration and invasion, at least partly via inhibition of VEGF-C. It was also demonstrated that the mRNA and protein expression levels of VEGF-C were frequently upregulated in ICC tissue and cells, and its expression level was inversely correlated with that of miR-101 in ICC tissue. In conclusion, the present study identified important roles for miR-101 and VEGF-C in ICC, suggesting that miR-101/VEGF-C signaling may be a promising diagnostic and/or therapeutic target for ICC. [ABSTRACT FROM AUTHOR]

MicroRNA-128 (miR-128) serves an important role in regulating growth, invasiveness, stem cell-like traits, differentiation and apoptosis of different types of tumor cells. Vascular endothelial growth factor-C (VEGF-C) has been associated with angiogenesis, lymphangiogenesis and regional lymph node metastasis and has previously been reported to have an anti-apoptotic and proliferative role in bladder cancer (BC). To investigate the regulation of miR-128 on VEGF-C expression and their effects on proliferation and metastasis of bladder cancer, T24 and 5637 BC cells were transfected with pre-miR-128, anti-miR-128 and their respective negative control. miR-128 was downregulated in BC tissues and cell lines, while the expression levels of VEGF-C were upregulated. The present results indicated that miR-128 negatively regulated VEGF-C expression in BC T24 and 5637 BC cells. VEGF-C is a direct target of miR-128 in BC cells. Overexpression of miR-128 inhibited cell proliferation, migration and invasion. Knockdown of miR-128 promoted proliferation, migration and invasion in BC cells. Therefore, downregulation mediated malignant progression of BC may be partly attributed to increased VEGF-C expression. Consequently, the findings of the present study provide a molecular basis for the role of miR-128/VEGF-C in the progression of human BC and indicate a novel target for treatment of BC. [ABSTRACT FROM AUTHOR]

Objective To explore the expression of vascular endothelial growth factor-C ( VEGF-C) in the occurrence and development of oral and maxillofacial lymphangioma (OML). Methods The tumor tissues (OML group) and the adjacent normal tissue (control group) from OML patients were collected. The real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of VEGF-C mRNA in the tissues. Immunohistochemistry (IHC) was used to detect the positive rate of VEGF-C protein expression in the tissues. ECV304 cells cultured in vitro were randomly divided into two groups: the observation group which was transfected with sh-VEGF-C plasmid and the control group transfected with control plasmid sh-control. After 48-hour transfection, qRT-PCR and Western blotting were used to detect the relative expression of VEGF-C protein, and MTT assay was used to detect the proliferation of the cells (absorbance value). Results In the OML group and control group, the expression of VEGF-C mRNA was 1.061 ±0.284 and 0.801 ±0.098, respectively; the positive expression rates of VEGF-C protein was 84. 10% (42/54) and 10. 13% (5/54) , respectively ( P <0. 05 or P < 0. 01 ). In the observation group and control group, the relative expression levels of VEGF-C mRNA were 0. 200 ±0. 058 and 0. 967 ±0. 017, respectively; the relative expression levels of VEGF-C protein were 0. 319 ±0. 021 and 0.965 ±0.039, respectively; the A values were 0. 393 ±0.015 and 0. 97 ±0. 004, respectively; significant differences were found between these two groups (all P <0.01 ). Conclusions VEGF-C is highly expressed in the OML tissues, and decreasing VEGF-C can inhibit the proliferation of human umbilical vein endothelial cells. [ABSTRACT FROM AUTHOR]

Vascular endothelial growth factor (VEGF)-C, as a lymphangiogenic factor, plays important roles in the progression of several malignancies. However, its clinical prognostic value in breast cancer still remains controversial. We performed a meta-analysis of available studies to assess the association between VEGF-C expression and the ou-tcomes of breast cancer patients.We searched eligible studies in three English databases (MEDLINE, EMBASE, and Web of Science) and two Chinese databases (Wanfang and Chinese National Knowledge Infrastructure databases). Key words used in the research included 'VEGF-C", "breast cancer", "immunohistochemistry", "breast neoplasma(s)", "breast carcinoma", "metastasis", and "prognosis". Fourteen studies with a total of 1, 573 breast cancer cases were finally included into the meta-analysis. The pooled odds ratios (ORs) with the corresponding 95% confidence interval (95% CIs) for lymph node metastasis, overall survival, and disease-free survival were calculated by using fixed-effects or random-effects models. Heterogeneity and publication bias were also assessed.Meta-analysis of random-effects model showed VEGF-C expression was associated with lymph node metastasis in patients with breast cancer (random-effects, OR = 2.14; 95 % CI 1.21-3.77, P = 0.009). VEGF-C expression was associated with poorer overall survival (fixed-effects, OR = 2.46, 95% CI: 1.46-4.14, P < 0.001) and disease-free survival (fixed-effects, OR = 2.10, 95% CI: 1.32-3.35, P = 0.002) in patients with breast cancer.VEGF-C expression is positively associated with lymph node metastasis in breast cancer, and VEGF-C detection in breast cancer might be an effective and feasible means to predict outcome.

The aim of the present study was to elucidate the associations between the expression of the vascular endothelial growth factor-C (VEGF-C)/VEGF receptor-3 (VEGFR-3) axis and lymphangiogenesis, regional lymph node metastasis and clinicopathological factors in oral tongue squamous cell carcinoma (OTSCC) using immunohistochemistry. The expression of VEGF-C, VEGFR-3 and podoplanin was immunohistochemically evaluated in specimens obtained from 65 patients with OTSCC (T1-2, N0) who had undergone radical surgery alone. The associations between the expression of VEGF-C, VEGFR-3 and podoplanin, and lymphangiogenesis, regional lymph node metastasis and clinocopathological factors were determined by immunohistochemical analysis. VEGF-C, VEGFR-3 and combined VEGF-C/VEGFR-3 expression was significantly higher in cases with regional recurrence compared with those without lymph node involvement (P<0.001). As regards lymphangiogenesis, a significant correlation was observed between podoplanin expression and VEGF-C, VEGFR-3 and combined VEGF-C/VEGFR-3 expression (P<0.001). Therefore, lymphangiogenesis in the peritumoral stroma was associated with lymph node metastasis. However, podoplanin expression did not exhibit a significant correlation with the progression of lymph node metastasis. The results of the present study suggest that the VEGF-C/VEGFR-3 axis may be associated with lymph node metastasis through lymphangiogenesis. Determining the VEGF-C/VEGFR-3 expression status may help predict which patients will develop regional recurrence and provide novel targets for therapies to suppress lymph node metastasis in the treatment of OTSCC. [ABSTRACT FROM AUTHOR]

Background/purpose Evidence shows that there is a relationship between hypoxia and inflammatory response in periodontitis. Hypoxia-inducible factor (HIF)-1α is a major regulator of energy homeostasis and cellular adaptation to low oxygen stress. Although experimental results demonstrate an association between HIF-1α and vascular endothelial growth factor (VEGF)-C in tumor angiogenesis, the role of HIF-1α and VEGF-C in the pathogenesis of periodontitis is still ambiguous. So far, limited attention has been given to the role of hypoxia and VEGF-C in periodontitis. The present study aimed to investigate the expression and distribution of HIF-1α and VEGF-C in gingival tissue samples from patients with different stages of chronic periodontitis and healthy individuals. Materials and methods A total of 56 samples were involved in this study, including moderate chronic periodontitis ( n = 20), advanced chronic periodontitis ( n = 20), and healthy control tissues ( n = 16). The gingival specimens were stained with hematoxylin and eosin for histopathology. The expression of HIF-1α and VEGF-C in gingival tissues was detected by immunohistochemical staining. Results HIF-1α and VEGF-C were found in gingival tissues from patients with different stages of chronic periodontitis as well as healthy control tissues. HIF-1α protein was expressed mainly in the epithelial layer of gingival tissues, and VEGF-C protein was mostly located in the connective tissue papilla of gingival tissues. Compared with healthy controls, the expression of HIF-1α and VEGF-C in chronic periodontitis groups was significantly higher (P < 0.01), and the density of HIF-1α and VEGF-C in advanced chronic periodontitis group was even significantly higher than that in the moderate chronic periodontitis group (P < 0.01). Conclusion Our results suggest that the expression of HIF-1α and VEGF-C increased with severity of periodontitis. So, we conclude that HIF-1α may play an important role in the pathophysiology of human periodontitis and may be related to the function of VEGF-C during periodontitis. [ABSTRACT FROM AUTHOR]

The aim of the current study was to elucidate the association between vascular endothelial growth factor C (VEGF-C) and resistance of bladder cancer cells to cisplatin and the underlying mechanism involving maspin. A total of 32 bladder cancer tissue samples from patients (18 males and 14 females with an average age of 65.9 years) were collected from the Fifth Affiliated Hospital of Zhengzhou University (Zhengzhou, China). All patients had undergone cisplatin-based combination chemotherapy. In addition, the BIU87 human bladder cancer cell line was cultured and a cisplatin-resistant subline (BIU87-CisR) was established by continuous exposure to cisplatin. The mRNA expression levels of VEGF-C and maspin in tissue samples, BIU87 cells and BIU87-CisR cells were analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Targeted inhibition of VEGF-C in BIU87-CisR cells was performed using small interfering (si)RNA technology and the alteration in levels of maspin was confirmed by RT-qPCR and western blot analysis. siRNA-treated and -untreated BIU87-CisR cells were divided into the following four groups: Control group (no drug treatment), 3 μM cisplatin treated group, 3 μM cisplatin + siRNA treated group and the siRNA treated group. Cell viability following treatment in each group was evaluated by the cell counting kit 8 assay. The cell cycle and apoptotic rate of BIU87-CisR cells was analyzed by propidium iodide (PI) staining and Annexin V-PI double staining with flow cytometry. Furthermore, pcDNA-maspin transfected BIU78-CisR cells were used to establish the effect of maspin on the sensitivity to cisplatin. VEGF-C expression in chemoresistant patients and BIU87-CisR cells was significantly increased compared with chemosensitive patients and normal BIU87 cells, respectively. By contrast, maspin levels were lower in chemoresistant patients and BIU87-CisR cells. Subsequent to VEGF-C inhibition, maspin expression was markedly increased. Cisplatin (3 μM) resulted in moderate proliferation inhibition of BIU87-CisR cells without siRNA pretreatment; however, significant inhibition was observed in the VEGF-C siRNA treated group. In addition, the cell cycle arrest and apoptosis induced by cisplatin was enhanced by VEGF-C inhibition. Overexpression of maspin was able to improve the sensitivity of BIU87-CisR cells to cisplatin. In conclusion, the resistance of bladder cancer cells to cisplatin may be induced by upregulation of VEGF-C, and inhibition of VEGF-C reverses resistance by elevating maspin expression levels. [ABSTRACT FROM AUTHOR]

The present study aimed to evaluate the expression of vascular endothelial growth factor C (VEGF-C) in human cholangiocarcinoma tissues and its role in metastasis in vitro. A total of 65 biopsy samples of cholangiocarcinoma, plus the FRH-0201 cell line, were investigated. The expression of VEGF-C in the human cholangiocarcinoma specimens was evaluated by immunohistochemistry (IHC). The effect of VEGF-C on tumor cell migration and proliferation was measured by MTT and Transwell assays in the FRH-0201 cell line. According to the IHC results, the biopsies of human cholangiocarcinoma were stained positively for VEGF-C, with a positive rate of 75.4% (49/65). Moreover, VEGF-C was expressed at a higher level in the patients with lymph node metastasis than in those without lymph node metastasis. In vitro, VEGF-C exhibited marked growth stimulation below the concentration of 5 ng/ml and was able to promote cholangiocarcinoma cell migration significantly. These findings suggested that VEGF-C may be a useful factor to predict lymph node metastasis in cholangiocarcinoma tissues and indicates that VEGF-C plays a significant role in proliferation and migration in cholangiocarcinoma cells. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of metformin combined with oxaliplatin on the expression levels of vascular endothelial growth factor-C (VEGF-C), vascular endothelial growth factor receptor-3 (VEGFR-3), CD44v6 and β-catenin of lung cancer xenograft in nude mice. Methods We established human lung cancer xenograft models in the nude mice.The nude mice were randomly divided into metformin-treated, oxaliplatin-treated, metformin combined with oxaliplatin-treated and untreated control groups. Forty-two days after administration, all the nude mice were sacrificed and tumor tissues were sampled. The protein and mRNA expression of VEGF-C, VEGFR-3, CD44v6 and β-catenin in the tumor tissues was detected by immumohistochemical method and florescent real-time quantitative PCR, repectively. Results The respective tumor inhibition rates of metformin-treated group , oxaliplatin-treated group and metformin combined with oxaliplatin-treated group were 28.97%, 34.37% and 50.27%. The expression levels of VEGF-C, VEGFR-3, CD44v6 and β-catenin protein and mRNA in the oxaliplatin-treated and metformin combined with oxaliplatin-treated groups were lower than those in the control group, respectively (all P<0.05). Compared with the metformin-treated group and oxaliplatin-treated group, the expression levels of VEGF-C, VEGFR-3, CD44v6 and β-catenin protein and mRNA of the metformin combined with oxaliplatin-treated group were lower, respectively (all P<0.05). F values about comparison of VEGF-C, VEGFR-3, CD44v6 and β-catenin proteins within four groups were 79.281, 151.275, 171.294 and 164.149, respectively (all P<0.01); F values about comparison of VEGF-C, VEGFR-3, CD44v6 andβ-catenin mRNA within four groups were 78.897, 81.029, 64.100 and 83.892, respectively (all P<0.01). Conclusions Using oxaliplatin or metformin can inhibit the expression of VEGF-C,VEGFR-3, CD44v6 and β-catenin. The combination of metformin and oxaliplatin can en-hance the anti-tumor efficacy. [ABSTRACT FROM AUTHOR]

Background: Tumor-induced lymphangiogenesis facilitates breast cancer progression by generating new lymphatic vessels that serve as conduits for tumor dissemination to lymph nodes and beyond. Given the recent evidence suggesting the implication of C-C chemokine ligand 21/chemokine receptor 7 (CCL21/CCR7) in lymph node metastasis, the aim of our study was to define the role of this chemokine pair in breast cancer-associated lymphangiogenesis. Methods: The expression analysis of CCL21/CCR7 pair and lymphatic endothelial cell (LEC) markers in breast cancer specimens was performed by means of quantitative real-time PCR. By utilizing CCR7 and CCL21 gene manipulated breast cancer cell implants into orthotopic sites of nude mice, lymphatic vessel formation was assessed through quantitative real-time PCR, immunohistochemistry and immunofluorescence assays. Finally, the lymphangiogenic potential of CCL21/CCR7 was assessed in vitro with primary LECs through separate functional assays, each attempting to mimic different stages of the lymphangiogenic process. Results: We found that CCR7 mRNA expression in human breast cancer tissues positively correlates with the expression of lymphatic endothelial markers LYVE-1, podoplanin, Prox-1, and vascular endothelial growth factor-C (VEGF-C). We demonstrated that the expression of CCL21/CCR7 by breast cancer cells has the ability to promote tumor-induced lymph-vascular recruitment in vivo. In vitro, CCL21/CCR7 chemokine axis regulates the expression and secretion of lymphangiogenic factor VEGF-C and thereby promotes proliferation, migration, as well as tube formation of the primary human LECs. Finally, we showed that protein kinase B (AKT) signaling pathway is the intracellular mechanism of CCR7-mediated VEGF-C secretion by human breast cancer cells. Conclusions: These results reveal that CCR7 and VEGF-C display a significant crosstalk and suggest a novel role of the CCL21/CCR7 chemokine axis in the promotion of breast cancer-induced lymphangiogenesis. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the effect of heat shock protein 90 (Hsp90)-specific inhibitor geldanamycin (GA) on the proliferation and apoptosis induced by vascular endothelial growth factor-C (VEGF-C) in cervical cancer cells. HeLa cells (1x106/ml) in the logarithmic growth phase were incubated without serum for 24 h. The cells were pretreated with kinase insert domain receptor antibody (KDR)-Ab (20 µg/ml), phosphatidylinositol 3-kinase (PI3K) i nhibitor LY294002 (3 µmol/l), mitogen-activated protein kinase (MAPK) inhibitor PD98059 (30 µmol/l) or Hsp90-specific inhibitor GA (10 µmol/l) for 30 min, and then treated with VEGF-C (50 ng/µl) for a further 24 h. The cells were harvested for MTT analysis, annexin V-FITC/propidium iodide double staining for early apoptosis and SDS-PAGE and western blot analysis in order to determine Hsp90, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and cyclin D1 expression. Treatment with VEGF-C alone induced Hsp90 protein expression in HeLa cells at all time-points. Hsp90 expression was increased 3.31-fold in VEGF-C treated HeLa cells, and this increase was attenuated in the treatment groups (2.17-, 1.69-, 1.82-fold in VEGF-C + KDR-Ab, VEGF-C + PD98059 and VEGF-C + LY294002, respectively). The proliferation of the VEGF-C-treated HeLa cells was increased ~2.13-fold, while that of the VEGF-C + GA-treated HeLa cells decreased 0.87-fold (P<0.05). Even low concentrations of GA (0.02 µmol/l) were found to inhibit the Bcl-2 and cyclin D1 protein expression induced by VEGF-C. Therefore, the results indicate that the Hsp90-specific inhibitor GA has a critical role in the proliferation and apoptosis induced by VEGF-C in cervical cancer cells. [ABSTRACT FROM AUTHOR]

Vascular endothelial growth factor ( VEGF)-C is an important lymphangiogenic factor involved in the lymphangiogenesis of gallbladder carcinoma ( GBC) and the lymph node metastasis of the tumor. Tumor necrosis factor ( TNF)-α, a key inflammatory cytokine responding to chronic inflammation of GBC, has been reported to stimulate the expression of VEGF-C in some nonneoplastic cells. But whether TNF-α promotes the expression of VEGF-C in GBC has yet to be determined. Therefore, in the present study, the concentration of TNF-α and VEGF-C and the lymphatic vessel density ( LVD) in the clinical GBC specimens were analyzed, and a linear correlation was found between the concentration of TNF-α and that of VEGF-C, the lymphatic vessel density ( LVD); The transcription and protein level of VEGF-C in NOZ cell line were detected by real-time polymerase chain reaction ( PCR) and enzyme linked immunosorbent assay ( ELISA), and TNF-α enhanced the expression of VEGF-C in NOZ cell lines in a dose and time-dependent manner. Lymphatic tube formation in vitro was observed in a three-dimensional coculture system consisting of HDLECs and NOZ cell lines, and lymphatic vessels of GBC in nude mice model was detected by immunohistochemistry. TNF-α promoted the tube formation of lymphatic endothelial cells in vitro and the lymphangiogenesis of GBC in nude mice; The nuclear factor ( NF)-κB binding site on the VEGF-C promoter was identified using Site-directed mutagenesis, electrophoretic mobility shift assay ( EMSA) and chromatin immunoprecipitation assay (Ch IP). Taken together, TNF-α can upregulate the expression of VEGF-C and promote the lymphangiogenesis of GBC via NF-κB combining with the promoter of VEGF-C. [ABSTRACT FROM AUTHOR]