Vascular Endothelial Growth factor-C mRNA Research Articles

Sodium accumulation in the skin is associated with higher density of skin lymphatic vessels in patients with arterial hypertension

PurposeRecent studies, conducted mainly on the rodent model, have demonstrated that regulatory pathway in the skin provided by glycosaminoglycans, nuclear factor of activated T cells 5 (NFAT5), vascular endothelial growth factor C (VEGF-C) and process of lymphangiogenesis may play an important role in extrarenal regulation of sodium (Na+) balance, body water volume, and blood pressure. We aimed to investigate the concentrations and relations among the main factors of this pathway in human skin to confirm that this regulatory axis also exists in humans. Patients and methodsSkin specimens from patients diagnosed with arterial hypertension and from control group were histologically and molecularly examined. ResultsThe primary hypertensive and control groups did not differ in Na+ ​concentrations in the skin. However, the patients with hypertension and higher skin Na+ concentration had significantly greater density of skin lymphatic vessels. Higher skin Na+concentration was associated with higher skin water content. In turn, skin water content correlated with factors associated with lymphangiogenesis, i.e. NFAT5, VEGF-C, and podoplanin (PDPN) mRNA expression in the skin. The strong mutual pairwise correlations of the expressions of NFAT5, VEGF-C, vascular endothelial growth factor D (VEGF-D) and PDPN mRNA were noted in the skin in all of the studied groups. ConclusionsOur study confirms that skin interstitium and the lymphatic system may be important players in the pathophysiology of arterial hypertension in humans. Based on the results of our study and existing literature in this field, we propose the hypothetical model which might explain the phenomenon of salt-sensitivity.

Uses of Vascular Endothelial Growth Factor C as a Lung Adenocarcinoma Prognostic Biomarker.

Lung adenocarcinoma (LUAD) is the most common type of lung cancer and a leading cause of death worldwide. Vascular endothelial growth factor C (VEGF-C) has been identified as a prognosis prediction marker for LUAD. However, VEGF-C protein expression does not appear to significantly relate to LUAD patient survival in several studies. We carried out a bioinformatic analysis to review the effect of VEGF-C mRNA expression on LUAD patient outcomes. GEPIA, UALCAN, TCGAportal, OncoLnc, LCE, GeneMANIA, Metascape, ImmuCellAI, and GSCA online databases were utilized. The expression levels of VEGF-C mRNA between normal tissue and LUAD tissue, overall survival (OS) analysis, function analysis, tumor microenvironment and drug sensitivity were conducted in the current study. We found that the expression level of VEGF-C mRNA was significantly lower in LUAD than normal tissue. Low expression of VEGF-C mRNA was also associated with better OS. VEGF-C expression was correlated with both NF1 and TP53 mutation status. No relationship was observed between VEGF-C and Tr1 or CD4 T-cell infiltrate scores. Additionally, VEGF-C was associated with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance. The sensitivity of 5-fluorouracil was positively correlated with VEGF-C, and the sensitivity of TGX221 was negatively correlated with VEGF-C. The activity of BI-2536 and BRD-A94377914 was positively correlated with VEGF-C. Novel LUAD prognostic biomarkers such as VEGF-C mRNA may aid diagnosis and treatment, and may help identify optimal LUAD populations for therapeutic treatments.

Vascular endothelial growth factor C gene expression and its serum level as potential biomarkers for obesity in Egyptian children

Background and objectivesObesity is a serious health issue among children, and it's linked to a higher risk of chronic illness. Increased adipose tissue, which is highly vascularized due to angiogenesis, causes adverse health problems. Angiogenesis and lymphangiogenesis are regulated by vascular endothelial growth factor C (VEGF-C), and poor lymphatic vessel function is linked to increased adipocyte accumulation. Therefore, our study analyzes VEGF-C gene expression and its serum level in children with obesity in Egypt and correlates them with metabolic, clinical, and biochemical parameters. Subjects & methodsWe compared VEGF-C gene expression in blood and its serum level from 50 unrelated children with obesity and 50 normal healthy controls chosen randomly of matched age and gender recruited from Pediatric Clinics, both inpatient and outpatient, Menoufia University, Egypt, from March to June 2021, in a case-control study. The expression levels of VEGF-C mRNA were evaluated using a real-time quantitative reverse transcription-polymerase chain reaction and the serum VEGF-C level was measured using an enzyme-linked immunosorbent assay (ELISA). ResultsFasting plasma glucose, fasting insulin, homeostatic model assessment for insulin resistance (HOMA-IR), triacylglycerol (TAG), total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDLC), serum VEGF-C, and VEGF-C expression levels were significantly higher in obese children group compared with the control (P < 0.001). In addition, serum VEGF-C levels were significantly inversely correlated with FT4 (P = 0.009) and significantly directly correlated with total cholesterol (P = 0.005) and LDL-C levels (P = 0.006) in the obese children group. Moreover, VEGF-C expression levels were also significantly inversely correlated with HDL-C levels (P = 0.001) and significantly directly correlated with fasting insulin (P = 0.002) and HOMA-IR (P = 0.001) in the obese children group. ConclusionVEGF-C gene expression and its serum levels could be used for follow-up of obese children with metabolic abnormalities.

Meningeal lymphatics regulate radiotherapy efficacy through modulating anti-tumor immunity

As a first-line treatment, radiotherapy (RT) is known to modulate the immune microenvironment of glioma, but it is unknown whether the meningeal lymphatic vessel (MLV)-cervical lymph node (CLN) network regulates the process or influences RT efficacy. Here, we show that the MLV-CLN network contributes to RT efficacy in brain tumors and mediates the RT-modulated anti-tumor immunity that is enhanced by vascular endothelial growth factor C (VEGF-C). Meningeal lymphatic dysfunction impaired tumor-derived dendritic cell (DC) trafficking and CD8+ T cell activation after RT, whereas tumors overexpressing VEGF-C with meningeal lymphatic expansion were highly sensitive to RT. Mechanistically, VEGF-C-driven modulation of RT-triggered anti-tumor immunity was attributed to C-C Motif Chemokine Ligand 21 (CCL21)-dependent DC trafficking and CD8+ T cell activation. Notably, delivery of VEGF-C mRNA significantly enhanced RT efficacy and anti-tumor immunity in brain tumors. These findings suggest an essential role of the MLV-CLN network in RT-triggered anti-tumor immunity, and highlight the potential of VEGF-C mRNA for brain tumor therapy.

Nucleoside-modified VEGFC mRNA induces organ-specific lymphatic growth and reverses experimental lymphedema

Lack or dysfunction of the lymphatics leads to secondary lymphedema formation that seriously reduces the function of the affected organs and results in degradation of quality of life. Currently, there is no definitive treatment option for lymphedema. Here, we utilized nucleoside-modified mRNA encapsulated in lipid nanoparticles (LNPs) encoding murine Vascular Endothelial Growth Factor C (VEGFC) to stimulate lymphatic growth and function and reduce experimental lymphedema in mouse models. We demonstrated that administration of a single low-dose of VEGFC mRNA-LNPs induced durable, organ-specific lymphatic growth and formation of a functional lymphatic network. Importantly, VEGFC mRNA-LNP treatment reversed experimental lymphedema by restoring lymphatic function without inducing any obvious adverse events. Collectively, we present a novel application of the nucleoside-modified mRNA-LNP platform, describe a model for identifying the organ-specific physiological and pathophysiological roles of the lymphatics, and propose an efficient and safe treatment option that may serve as a novel therapeutic tool to reduce lymphedema.

Long Noncoding RNA ASLNC07322 Functions in VEGF-C Expression Regulated by Smad4 during Colon Cancer Metastasis

Deletion and mutation of the Smad4 gene are favorable events for the progression of colon cancer, which is related to the negative regulation of vascular endothelial growth factor C (VEGF-C). However, the regulatory mechanism between Smad4 and VEGF-C remains unclear. We reported first that Smad4 can increase the transcription of miR-128-3p, a microRNA targeting VEGF-C mRNA, resulting in a negative correlation between Smad4 and VEGF-C. Moreover, we found that Smad4 combined with Smad3 can positively regulate VEGF-C during colon cancer metastasis through binding to VEGF-C gene promoter. Further, results revealed a mechanism that long noncoding RNA (lncRNA) ASLNC07322 increased specifically in metastatic colon cancer and decreased miR-128-3p as a sponge, leading to a subsequent elevation of VEGF-C. In a word, there are two pathways in the progression of colon cancer, including Smad4/miR-128-3p/VEGF-C and Smad4/VEGF-C pathways in non-metastatic and metastatic colon cancer, respectively. ASLNC07322 crucially controlled this negative and positive regulatory transformation between them. Additionally, ASLNC07322 knockdown combined with Smad4 overexpression could efficiently inhibit lymphatic endothelial cells (LECs) proliferation and tube formation in vitro, as well as tumor growth and lymphangiogenesis in vivo. These data explained the underlying mechanism of Smad4 contribution on VEGF-C expression during metastasis where ASLNC07322 functions vitally as a switch in colon cancer.

Effect of oral mucosal transplantation on the expression of EGF and VEGF-C during skin wound repair.

This study evaluated the effects of oral mucosal transplantation on epidermal growth factor (EGF) and vascular endothelial growth factor C (VEGF-C) in skin wound repair. Sixty-four rats were randomly separated into group A, B, C and D (16 rats in each group). The right abdomen skin was excised 1, 3, 5 and 7 days after injury, respectively. Oral mucosa of the rat tongue was transplanted to the right abdomen skin. Fourteen days after the healing of the oral mucosa graft, the rat skin full-thickness model was prepared at the transplant site (the study group) and the contralateral site (the control group). Rats in each group were anesthetized and sacrificed at 1, 3, 5 and 7 days after injury. Expression of EGF and VEGF-C in skin tissue was detected by RT-qPCR and ELISA. At 3 days, expression levels of EGF and VEGF-C mRNA and protein in skin tissue were significantly higher than those at 1 day (P<0.05). At 5 days, expression levels of EGF and VEGF-C mRNA and protein in skin tissue were significantly higher than those at 3 days (P<0.05). At 7 days, expression levels of EGF mRNA and protein in skin tissue were significantly lower than those at 5 days (P<0.05), while VEGF-C levels were significantly increased (P<0.05). Expression levels of EGF and VEGF-C mRNA and protein in the skin tissue of the study group were significantly lower than those in the control group at all days (P<0.05). EGF and VEGF-C may be involved in scar formation, and play an important role in the process of skin wound repair.

VEGFC acts as a double-edged sword in renal cell carcinoma aggressiveness.

Hypoxic zones are common features of metastatic tumors. Due to inactivation of the von Hippel-Lindau gene (VHL), renal cell carcinomas (RCC) show constitutive stabilization of the alpha subunit of the hypoxia-inducible factor (HIF). Thus, RCC represents a model of chronic hypoxia. Development of the lymphatic network is dependent on vascular endothelial growth factor C (VEGFC) and lies at the front line of metastatic spreading. Here, we addressed the role of VEGFC in RCC aggressiveness and the regulation of its expression in hypoxia.Methods: Transcriptional and post transcriptional regulation of VEGFC expression was evaluated by qPCR and with reporter genes. The involvement of HIF was evaluated using a siRNA approach. Experimental RCC were performed with immuno-competent/deficient mice using human and mouse cells knocked-out for the VEGFC gene by a CRISPR/Cas9 method. The VEGFC axis was analyzed with an online available data base (TCGA) and using an independent cohort of patients.Results: Hypoxia induced VEGFC protein expression but down-regulated VEGFC gene transcription and mRNA stability. Increased proliferation, migration, over-activation of the AKT signaling pathway and enhanced expression of mesenchymal markers characterized VEGFC-/- cells. VEGFC-/- cells did not form tumors in immuno-deficient mice but developed aggressive tumors in immuno-competent mice. These tumors showed down-regulation of markers of activated lymphocytes and M1 macrophages, and up-regulation of M2 macrophages markers and programmed death ligand 1 (PDL1). Over-expression of lymphangiogenic genes including VEGFC was linked to increased disease-free and overall survival in patients with non-metastatic tumors, whereas its over-expression correlated with decreased progression-free and overall survival of metastatic patients.Conclusion: Our study revisited the admitted dogma linking VEGFC to tumor aggressiveness. We conclude that targeting VEGFC for therapy must be considered with caution.

Shikonin Suppresses Lymphangiogenesis via NF-κB/HIF-1α Axis Inhibition.

Lymphangiogenesis, the formation of lymphatic vessels from preexisting ones, promotes cancer growth and metastasis. Finding natural compounds with anti-lymphangiogenic activity will be useful for preventive treatment of lymphatic metastasis. Shikonin, an ingredient of a traditional Japanese and Chinese medicinal herb Lithospermum erythrorhizon, has been widely used in several pharmaceutical and cosmetic preparations, as well as in food colorants. Shikonin has been reported to inhibit lymphangiogenesis in vitro, but the mechanism of inhibition has not been determined. The aim of this study is to investigate the mechanism of anti-lymphangiogenesis of shikonin in primary human lymphatic endothelial cells (HMVEC-dLy). Shikonin, at non-toxic concentrations, significantly inhibited cord formation ability of lymphatic endothelial cells in a dose- and time-dependent manner. Western blotting analysis showed that shikonin decreased nuclear factor-kappaB (NF-κB) activation, as indicated by phosphorylation and nuclear translocation of NF-κB p65, and also reduced both mRNA and protein levels of hypoxia-inducible factor-1 (HIF-1)α. Use of an NF-κB inhibitor (BAY 11-7085) and HIF-1α small interfering RNA (siRNA) transfection revealed that NF-κB activation was upstream of HIF-1α expression, which controls cord formation by HMVEC-dLy. In addition, the reduction of vascular endothelial growth factor C (VEGF-C) and vascular endothelial growth factor receptor-3 (VEGFR-3) mRNA levels were also found in HMVEC-dLy that treated with shikonin. In conclusion, shikonin inhibits lymphangiogenesis in vitro by interfering the NF-κB/HIF-1α pathway and involves in suppression of VEGF-C and VEGFR-3 mRNA expression.

RIP1 regulates TNF-α-mediated lymphangiogenesis and lymphatic metastasis in gallbladder cancer by modulating the NF-κB-VEGF-C pathway.

BackgroundTumor necrosis factor alpha (TNF-α) enhances lymphangiogenesis in gallbladder carcinoma (GBC) via activation of nuclear factor (NF-κB)-dependent vascular endothelial growth factor-C (VEGF-C). Receptor-interacting protein 1 (RIP1) is a multifunctional protein in the TNF-α signaling pathway and is highly expressed in GBC. However, whether RIP1 participates in the signaling pathway of TNF-α-mediated VEGF-C expression that enhances lymphangiogenesis in GBC remains unclear.MethodsThe RIP1 protein levels in the GBC-SD and NOZ cells upon stimulation with increasing concentrations of TNF-α as indicated was examined using Western blot. Lentiviral RIP1 shRNA and siIκBα were constructed and transduced respectively them into NOZ and GBC-SD cells, and then PcDNA3.1-RIP1 vectors was transduced into siRIP1 cell lines to reverse RIP1 expression. The protein expression of RIP1, inhibitor of NF-κB alpha (IκBα), p-IκBα, TAK1, NF-κB essential modulator were examined through immunoblotting or immunoprecipitation. Moreover, VEGF-C mRNA levels were measured by quantitative real-time polymerase chain reaction, VEGF-C protein levels were measured by immunoblotting and enzyme-linked immunosorbent assay, and VEGF-C promoter and NF-κB activities were quantified using a dual luciferase reporter assay. The association of NF-κB with the VEGF-C promoter was analysed by chromatin immunoprecipitation assay. A three-dimensional coculture method and orthotopic transplantation nude mice model were used to evaluate lymphatic tube-forming and metastasis ability in GBC cells. The expression of RIP1 protein, TNF-α protein and lymphatic vessels in human GBC tissues was examined by immunohistochemistry, and the dependence between RIP1 protein with TNF-α protein and lymphatic vessel density was analysed.ResultsTNF-α dose- and time-dependently increased RIP1 protein expression in the GBC-SD and NOZ cells of GBC, and the strongest effect was observed with a concentration of 50 ng/ml. RIP1 is fundamental for TNF-α-mediated NF-κB activation in GBC cells and can regulate TNF-α-mediated VEGF-C expression at the protein and transcriptional levels through the NF-κB pathway. RIP1 can regulate TNF-α-mediated lymphatic tube formation and metastasis in GBC cells both in vitro and vivo. The average optical density of RIP1 was linearly related to that of TNF-α protein and the lymphatic vessel density in GBC tissues.ConclusionWe conclude that RIP1 regulates TNF-α-mediated lymphangiogenesis and lymph node metastasis in GBC by modulating the NF-κB-VEGF-C pathway.

Effect of High VEGF-C mRNA Expression on Achievement of Complete Remission in Adult Acute Myeloid Leukemia

Although vascular endothelial growth factor-C (VEGF-C) is known to be expressed in acute myeloid leukemia (AML) blasts, the relevance of VEGF-C in the clinical setting remains to be fully explored. We examined the effect of VEGF-C on achievement of complete remission (CR) in adult de novo AML and immune cell population profiles according to VEGF-C mRNA expression. In comparison of VEGF-C expression between the no-CR and CR groups, the CR group showed a trend toward higher levels of plasma VEGF-C (P = .088), whereas mRNA expression of VEGF-C was downregulated (P = .008). Next, patients with continuous data for VEGF-C were divided into two groups (low vs. high) by a ROC curve analysis. The low- versus high-level groups for plasma VEGF-C (RR of 0.20, P = .030), mRNA expression of VEGF-C (RR of 18.75, P = .003), and the ratio of plasma level to mRNA expression (RR of 0.05, P = .007) were potential predictors of CR on univariate analysis. After adjusting for potential clinical factors including genetic group, multivariate analyses revealed that high VEGF-C mRNA expression was an independent risk factor for failure of induction chemotherapy. Furthermore, patients with high VEGF-C mRNA expression had a lower frequency of NKT and CD8+ cells and showed a trend for a lower frequency of NK cells. These results suggest that interruption of VEGF-C signaling might be a potential therapeutic target for antileukemic treatment in AML patients.

MicroRNA-186 regulates the invasion and metastasis of bladder cancer via vascular endothelial growth factor C.

The present study aimed to investigate the expression of microRNA (miRNA or miR)-186 in tumor tissue, blood and urine from patients with bladder cancer. The mechanism by which miR-186 regulates the invasion and metastasis of bladder cancer was also assessed. A total of 76 patients who underwent surgical resection of bladder cancer tissues between August 2012 and January 2016 were included in the present study. Blood and urine samples were also collected from the 76 patients and another 66 healthy subjects. Expression of vascular endothelial growth factor C (VEGF-C) mRNA and miR-186 was measured using reverse transcription-quantitative polymerase chain reaction. Western blot analysis was performed to assess VEGF-C protein expression in tumor tissues. The content of VEGF-C protein in blood and urine samples was measured using an enzyme-linked immunosorbent assay. To identify the direct interaction between miR-186 and VEGF-C mRNA, a dual luciferase reporter assay was performed. The present findings demonstrated that VEGF-C mRNA expression in tumor tissues, blood and urine of bladder cancer patients was upregulated. VEGF-C protein expression in bladder cancer tissues was also enhanced. VEGF-C protein content in blood and urine from bladder cancer patients was elevated, consistent with the results for VEGF-C mRNA. Expression of miR-186 was reduced in tumor tissues, blood and urine. Dual luciferase reporter assay demonstrated that miR-186 regulated the expression of VEGF-C by binding with its 3'-untranslated region. Therefore, the results of the present study indicate that the expression of VEGF-C mRNA and protein is upregulated in tumor tissues, blood and urine from patients with bladder cancer, while that of miR-186 is downregulated in these samples. miR-186 potentially regulates the invasion and metastasis of bladder cancer via VEGF-C, and may become a gene marker for bladder cancer in the future.

Clinical Impact of Different Levels of VEGF-C on Leukemic Blasts in the Bone Marrow and Peripheral Blood of Acute Myeloid Leukemia

Background: Vascular endothelial growth factor-C (VEGF-C) is a lymph-angiogenic growth factor and in general, transmits intracellular signals, resulting in cell proliferation and survival. Previous studies have reported that VEGF-C is important for cancer progression based on the autocrine VEGF-C loop promoting the invasion and metastasis of cancer cells as well as the spread of cancer cells by active recruitment of new lymphatics by tumor-derived VEGF-C. Acute myeloid leukemia (AML) blasts express VEGF-C and its receptors. A few studies demonstrated that high VEGF-C mRNA expression of AML blasts were related to increased in vitro and in vivo drug resistance and could predict adverse long-term outcomes. However, whether the expression of VEGF-C in the bone marrow (BM) and peripheral blood (PB) has a similar role in the pathophysiology of AML remains unclear.Methods: In this study, we analyzed plasma levels of VEGF-C in both BM and PB samples of AML patients. The levels of VEGF-C were measured using a commercially available ELISA in the newly diagnosed AML patients (58 in BM, 26 in PB), patients in complete remission (CR) (26 in BM, 20 in PB), and refractory/relapsed AML (15 in BM, 10 in PB). In addition, response after 1st induction chemotherapy was assessed in 67 evaluable patients and to create the predictive model for an achievement of CR, logistic regression was used after log transformation of VEGF-C levels.Results: In the BM of patients with newly diagnosed AML, the level of VEGF-C was 47.87 ± 12.4 pg/ml which was significantly lower than that of refractory/relapsed AML [518.3 ± 320.0 pg/ml (P=0.005)] but there was no difference, compared to that of patients in CR [44.57 ± 8.44 pg/ml (P = 0.865)]. In contrast no trend was observed in the PB samples. Next, to create the predictive model for an achievement of CR, sixty-seven evaluable patients who received standard induction chemotherapy were analyzed. Thirty-seven men and 30 women were included. With a median age of 49 years (range, 20-78), the distribution of favorable, intermediate-I, intermediate-II, and adverse cytogenetic risk (ELN) were 25%, 31%, 21%, and 22%, respectively. The patients with continuous values of Log10VEGF-C were divided into 2 groups (low vs. high levels) by a ROC curve analysis. Univariate analysis showed that high levels in BM samples were associated with achievement of CR after 1st induction chemotherapy. Ultimately, multivariate analyses revealed that low levels of Log10VEGF-C showed a trend for failure to achieve a response of CR (RR of 0.24, P = 0.065) and intermediate II/adverse cytogenetic risk was associated with a failure of CR, compared to favorable/intermediate I (RR of 0.21, P = 0.036). In contrast, in the PB samples, a low value of Log10VEGF-C was an independent factor for achievement of CR (RR of 28.7, P = 0.043).Conclusion: Our data demonstrate that the high VEGF-C level in the BM samples at diagnosis was associated with a trend toward higher CR rate and high VEGF-C level in the PB samples was significantly related to a failure of CR, suggesting discrepancy of the role of VEGF-C level in the BM and PB. Further studies on the different mechanism of the VEGF-C/VEGF-receptor pathway in the BM and PB are warranted. DisclosuresNo relevant conflicts of interest to declare.

Interleukin-6 Induces Vascular Endothelial Growth Factor-C Expression via Src-FAK-STAT3 Signaling in Lymphatic Endothelial Cells.

Elevated serum interleukin-6 (IL-6) levels correlates with tumor grade and poor prognosis in cancer patients. IL-6 has been shown to promote tumor lymphangiogenesis through vascular endothelial growth factor-C (VEGF-C) induction in tumor cells. We recently showed that IL-6 also induced VEGF-C expression in lymphatic endothelial cells (LECs). However, the signaling mechanisms involved in IL-6-induces VEGF-C induction in LECs remain incompletely understood. In this study, we explored the causal role of focal adhesion kinase (FAK) in inducing VEGF-C expression in IL-6-stimulated murine LECs (SV-LECs). FAK signaling blockade by NSC 667249 (a FAK inhibitor) attenuated IL-6-induced VEGF-C expression and VEGF-C promoter-luciferase activities. IL-6’s enhancing effects of increasing FAK, ERK1/2, p38MAPK, C/EBPβ, p65 and STAT3 phosphorylation as well as C/EBPβ-, κB- and STAT3-luciferase activities were reduced in the presence of NSC 667249. STAT3 knockdown by STAT3 siRNA abrogated IL-6’s actions in elevating VEGF-C mRNA and protein levels. Moreover, Src-FAK signaling blockade reduced IL-6’s enhancing effects of increasing STAT3 binding to the VEGF-C promoter region, cell migration and endothelial tube formation of SV-LECs. Together these results suggest that IL-6 increases VEGF-C induction and lymphangiogenesis may involve, at least in part, Src-FAK-STAT3 cascade in LECs.

The relationship among the expressions of vascular endothelial growth factor-C and its receptor and the cervical cancer growth and lymph node metastasis

Objective To study the relationship between the expressions of vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor receptor-2 (KDR) in cervical carcinoma and the formation of cervical cancer and lymph node metastasis. Methods We selected 72 cervical carcinoma tissues, their corresponding adjacent tissues and 36 normal cervical tissues which have been resected in the Maternal and Child Health Care Hospital of Baoji of Shaanxi Province from January 2010 to December 2013. The mRNA and protein expressions of VEGF-C and KDR were examined by semi-quantitative PCR and enzyme linked immunosorbent assay in these tissues. The relationships between the expressions of VEGF-C and KDR and the formation of cervical cancer and lymph node metastasis were analyzed. Results The mRNA levels of VEGF-C in 72 cases of cervical cancer tissues and its corresponding adjacent tissues were 4.67±1.05 and 2.02±0.65, which were significantly higher than those in normal cervical tissues (0.36±0.06), with significant differences (t=2.247, P=0.025; t=1.379, P=0.027). The protein levels of VEGF-C in 72 cases of cervical cancer tissues and their corresponding adjacent tissues were 68.30±17.10 and 48.20±12.70, which were significantly higher than those in normal cervical tissues (18.40±10.70), with significant differences (t=4.357, P=0.016; t=6.337, P=0.012). The mRNA levels of KDR in 72 cases of cervical cancer tissues and their corresponding adjacent tissues were 3.52±0.95 and 1.92±0.87, which were significantly higher than those in normal cervical tissues (0.72±0.36), with significant differences (t=3.127, P=0.023; t=1.214, P=0.028). The protein levels of KDR in 72 cases of cervical cancer tissues and their corresponding adjacent tissues were 47.20±15.60 and 38.60±11.30, which were significantly higher than those in normal cervical tissues (16.40±9.40), with significant differences (t=3.667, P=0.020; t=0.986, P=0.032). The expression level of VEGF-C protein in 72 cases of cervical cancer tissues was not correlated with age (χ2=0.54, P=0.17), tissue type (χ2=0.34, P=0.25), depth of invasion (χ2=5.39, P=0.08), pathological grade (χ2=0.78, P=0.11), but was correlated with tumor size (χ2=22.34, P=0.02), clinical stage (χ2=32.14, P=0.01) and lymph node metastasis (χ2=15.58, P=0.03). The expression level of its receptor KDR was correlated with tumor size (χ2=13.78, P=0.04), tissue type (χ2=32.74, P=0.01), pathological grade (χ2=13.72, P=0.04), depth of invasion (χ2=10.27, P=0.04), clinical staging (χ2=20.25, P=0.02) and lymph node metastasis (χ2=19.52, P=0.02), but was not correlated with age (χ2=4.17, P=0.09). Conclusion The expression levels of VEGF-C and KDR are correlated with the growth, invasion and metastasis of cervical cancer, which are good indicators of the lymph node metastasis. Key words: Uterine cervical neoplasms; Lymphatic metastasis; Vascular endothelial growth factor C

Effects of curcumin combined with cisplatin on growth and lymphatic metastasis of cervical cancer xeno-grafts in nude mice

Objective To analyze the effects of curcumin combined with cisplatin on the growth and lymphatic metastasis of cervical cancer xenografts in nude mice. Methods The human cervical carcinoma Caski cells xenotransplanted tumor models were established. Inoculated mice were randomly divided into 4 groups according to random number table: control (normal saline 10 ml/kg), Cur (curcumin 100 mg/kg), Cis (cisplatin 3 mg/kg) and Cur+ Cis group (100 mg/kg curcumin+ 3 mg/kg cisplatin). The tumor volume and anti-tumor rate were calculated. The mRNA levels of macrophage migration inhibitory factor (MIF) and vascular endothelial growth factor-C (VEGF-C) were analyzed by real-time fluorescent quantitative polymerase chain reaction (PCR). Results The inhibitory rates of Cur group, Cis group and Cur+ Cis group were 40.8%, 53.3% and 60.0%. After 15 days treatment, the tumor volumes of the control group, Cur group, Cis group and Cur+ Cis group were (123.44±35.62), (71.72±28.36), (65.47±18.32), (53.44±10.79)mm3. The growth rates of tumor volume in Cur group, Cis group and Cur+ Cis group were slower, Cur+ Cis group could more effectively inhibit tumor volume growth. The difference among the four groups had statistically signi-ficance (F=16.890, P=0.000). Real-time fluorescent quantitative PCR detection showed that compared with the control group (1.000), the MIF mRNA expressions in Cur group, Cis group and Cur+ Cis group were 0.322±0.094, 0.154±0.006 and 0.136±0.007, VEGF-C mRNA expressions in the three groups were 0.312±0.068, 0.263±0.072 and 0.221±0.041. The differences among groups had statistically significance (F=220.279, P=0.000; F=143.250, P=0.000). MIF mRNA was positively related with VEGF-C mRNA (r=0.815, P=0.001). Conclusion Curcumin combined with cisplatin can inhibit the growth of Carski cell xenografts in nude mice. Through the down-regulating expression of MIF mRNA and VEGF-C mRNA, cisplatin and curcumin can inhibit the lymphatic metastasis of cervical cancer, and it may be one of the important mechanisms of its anti-tumor effects. Key words: Curcumin; Uterine cervical neoplasms; Cisplatin; Vascular endothelial growth factor C; Macrophage migration inhibitory factor

Elevated Expression of VEGF-C and Its Receptors, VEGFR-2 and VEGFR-3, in Patients with Mesial Temporal Lobe Epilepsy.

Mesial temporal lobe epilepsy (MTLE) is a frequent form of focal intractable epilepsy in adults. We previously reported overexpression of vascular endothelial growth factor C (VEGF-C) and its receptors, VEGFR-2 and VEGFR-3, in epilepsy-associated tuberous sclerosis complex. To identify whether VEGF-C and its receptors are involved in epileptogenesis of MTLE, we investigated the levels and expression pattern of VEGF-C and its receptors in temporal neocortex and hippocampus (HPC) from 28 patients with MTLE and ten control (CTX) subjects. Real-time quantitative polymerase chain reaction and Western blotting results revealed upregulated mRNA and immunoreactive protein levels of VEGF-C, VEGFR-2, and VEGFR-3 in the MTLE group compared to the control groups. Immunohistochemistry and double-labeled immunofluorescence showed that VEGF-C was highly expressed in neurons and astrocytes, including reactive astrocytes and vascular endothelial cells, VEGFR-2 was expressed at a high level in reactive astrocytes and vascular endothelial cells, but not in neurons, whereas VEGFR-3 was only overexpressed in reactive astrocytes. Taken together, these findings suggest that VEGF-C and its receptors, VEGFR-2 and VEGFR-3, may contribute to the epileptogenesis of MTLE.

Heparanase regulates in vitro VEGF-C expression and its clinical significance to pancreatic ductal cell adenocarcinoma.

Heparanase (HPSE) and vascular endothelial growth factor C (VEGF-C) are important cytokines that promote metastasis and angiogenesis in numerous malignant neoplasms, however, their association remains unclear in pancreatic ductal cell adenocarcinoma (PDAC). The present study aimed to investigate whether HPSE has a positive correlation with VEGF-C expression and to uncover the role it plays in the in vitro invasion of BxPC-3 cells (a pancreatic carcinoma cell line), and to analyze the value of joint detection of HPSE and VEGF-C for PDAC patients. A recombinant plasmid, GV230/HPSE was constructed and BxPC-3 cells were transiently transfected with GV230/HPSE or siRNA against HPSE. The expression levels of HPSE and VEGF-C were compared using reverse transcription quantitative PCR (RT-qPCR) and immunoblotting. The metastatic potential of treated BxPC-3 cells was evaluated using a Transwell® invasion assay. The relative mRNA levels of HPSE and VEGF-C in 34 PDAC specimens were assessed by RT-qPCR. The results of the RT-qPCR demonstrated a 10.7- and 3.24-fold elevation (P<0.01) of HPSE mRNA and VEGF-C mRNA, respectively, in GV230/HPSE group, whereas the HPSE siRNA group were downregulated for these mRNAs (−2.45-fold, P<0.01; −1.84-fold, P<0.01). The same pattern for protein expression was detected using immunoblot assays. In Transwell® invasion assays 138±5 cells in GV230/HPSE group and 53±4 cells in siRNA group migrated through the Matrigel®. A negative correlation between the mRNA levels of HPSE and VEGF-C in PDAC specimens and the prognosis factors of the postoperative patients was identified. Spearman rank correlation analysis indicated a positive correlation between HPSE and VEGF-C in PDAC (r=0.812, P<0.01). HPSE regulates the expression of VEGF-C and facilitates invasion of BxPC-3 in vitro. Joint detection of HPSE and VEGF-C may therefore be clinically useful in determining the prognosis of pancreatic cancer patients.

Influence of Photodynamic Therapy on Apoptosis and Invasion of Human Cholangiocarcinoma QBC939 Cell Line.

To investigate the effect of photodynamic therapy (PDT) mediated by hematoporphyrin derivative (HPD) on apoptosis and invasion of cholangiocarcinoma QBC939 cell lines. In vitro cultured cholangiocarcinoma QBC939 cell line was exposed to 2, 4, 6, 8, 10, 12, and 14 μg/ml HPD with 5, 10, and 15 J/cm2 light intensity, respectively. The optical density at 450 nm of the QBC939 cells was measured by CCK8 assay and its growth inhibition ratio was calculated. Flow cytometry and transwell migration assay were applied to detect cell apoptosis and invasion respectively. RT-PCR and immunocytochemistry analyses were used to detect expressions of vascular endothelial growth factor-C (VEGF-C), cyclooxygenase-2 (COX-2), and proliferating cell nuclear antigen (PCNA). Enzyme-linked immunosorbent assay (ELISA) was carried out to examine the secretion of VEGF-C and COX-2 in QBC939 cells. Exposure to HPD-PDT can significantly suppress the growth of QBC939 cells (all P<0.05). HPD-PDT can promote apoptosis of QBC939 cells at the early stage. When the concentration of HPD was 2 μg/ml and light irradiation was 5 J/cm2, HPD-PDT had no obvious inhibitory effect on QBC939 cell growth, but can obviously inhibit cell invasion, and significant difference was observed between the HPD-PDT and control groups (P<0.01). The HPD-PDT can reduce the mRNA and protein expressions of VEGF-C, COX-2, and PCNA, and decrease the secretion of VEGF-C and COX-2 in QBC939 cells. PDT could promote apoptosis and inhibit growth and invasion of cholangiocarcinoma cells QBC939 in vitro.

Tumor-induced VEGF-C overexpression in retroperitoneal lymph nodes in VX2 carcinoma-bearing rabbits.

ObjectiveTo establish the retroperitoneal lymph node (RLN) metastasis model of cervical carcinoma in rabbits and evaluate the relationship of vascular endothelial growth factor-C (VEGF-C) expression and the lymph node status.MethodsForty-eight rabbits were injected with VX2 cells or RPMI solution at muscular mucosae of the myometrium 0.5 cm away from the cervix. Animals were treated with or without cis-diamminedichloroplatinum(II) (cisplatin: DDP) and sacrificed on days 15, 21, and 27 post-VX2 or RPMI injections. Tumor mass and RLNs were examined histopathologically. Quantitative real-time PCR was used to examine the changes in VEGF-C mRNA expression. Levels of VEGF-C protein expression in tissues were determined using immunohistochemistry staining.ResultsDevelopment of VX2 cervical carcinoma and the RLNs metastasis was confirmed with pathological examination. Significantly increased tumor volume was observed on days 15, 21, and 27 postinjection (P<0.05). The enlargement of RLNs was found on day 21. Expression of VEGF-C was significantly upregulated in peripheral white blood cells, tumor mass, and RLNs in an association with cancer progression. DDP resulted in a suppression of VEGF-C expression, whereas the influences on tumor mass and lymphatic metastasis were insignificant.ConclusionElevated VEGF-C expressions in peripheral white blood cells and RLNs are associated with tumor progression and lymphatic metastasis. DDP treatment inhibits VEGF-C expression and fails to protect against metastatic cervical cancer.