Investigation the Relationship among VEGF-C and Its Receptor VEGFR-3/Flt-4 in Tumour Lymphagiogenesis, Proliferation and Metastasis of Human Carcinoma
|School||Huazhong University of Science and Technology|
|Course||Obstetrics and Gynaecology|
|Keywords||vascular endothelial growth factor-C vascular endothelial growth factor receptor-3 cyclooxygenase-2 human cervical carcinoma cell line function lymphagiogenesis lymph node metastasis cervical cancer VEGF-C Cell proliferation|
Objective: Studies have suggested that the vascular endothelial growth factor-C and its receptor VEGFR-3/FLT-4 play an important role in tumour growth and metastasis. We conducted the present study to clarify whether human cervical cancer cells express factor-C and VEGFR-3/FLT-4 and their biological significance. Another aim of the study was to investigate the relationship between VEGF-C and cyclooxygenase-2 (COX-2) in human cervical cancer cells.Methods: We examined expression of VEGF-C and VEGFR-3 in four human cervical cancer cell lines Hela, Siha, C33a and Caski by flow cytometer, Western blot and RT-PCR, investigated the Functions of VEGF-C and VEGFR-3. We also examined the expression of COX-2 by cell immunochemical and investigated the relationship between the expression of VEGF-C and COX-2. Further, we evaluated whether recombinant human VEGF-C (rhVEGF-C) and COX-2-specific inhibitor NS398 may affect the proliferation and migration of human cervical cancer cells.Results: VEGF-C and VEGFR-3 proteins and mRNA were all assaied in the Four human Cervical carcinoma cell lines. But there were Differences among the expression of them. Usually, The level of VEGF-C expression is very low(<30%) while VEGFR-3 is very high(≥90%). COX-2 expressed in the cytoplasm of cervical carcinoma cells and the lever of COX-2 expression was coincidence with the expression of VEGF-C. Furthermore, VEGF-C proteins and mRNA of Hela cells significantly increased when stimulated with rhVEGF-C or decreased when treated with NS398. However, VEGFR-3 expression did not change under the same circumstance. rhVEGF-C or NS398 induced a significant increase or decrease of the proliferate activity of Hela cells in a dose-dependent manner(P < 0.05). The optimal concentration of rhVEGF-C and NS398 was 375 ng/mL and 60μg/mL, respectively. Transwell assay showed that the invasive ability in rhVEGF-C was enhanced (55.76±2.16) (P<0.01), while attenuated in NS398 (25.45±3.67) (P<0.05) or in monoclonal anti-human VEGFR-2 antibody+ the blocking peptide of VEGFR-3 (27.15±3.99) (P<0.05) as compared with those in control(40.38±0.72).Conclusions: The results indicated that VEGF-C, VEGFR-3 and COX-2 were all expressed in human cervical carcinoma cells and the expression of COX-2 was related to the expression of VEGF-C. VEGF-C and its receptors may play a crucial role in the regulation of tumor growth and metastasis. COX-2 may be a regulator of VEGF-C expression and its specific inhibitor NS398 may have therapeutic effect on the inhibition of VEGF-C mediated tumor growth and metastases in human cervical carcinoma cell lines. Objective: lymphatic vascular invasion and lymph node metastasis was shown to play a key role in the progression of cervical cancer. Recent studies in different tumors have shown that vascular endothelial growth factor-C (VEGF-C), a specific growth factor of lymphatics, is involved in tumor-induced lymphagiogenesis and lymphatic metastatic spread. The purpose of the present study was to investigate the expression of VEGF-C and its receptor VEGFR-3 in human cervical cancer to elucidate its role in tumor progression, lymphagiogenesis and lymph node metastasis. Another aim of the study was to investigate the relation between VEGF-C and cyclooxygenase-2 (COX-2) in cervical cancer.Methods: The expression of VEGF-C, VEGFR-3 and COX-2 was evaluated in 78 cases of surgical speciments of cervical diseases by immunohistochemical staining. Clinical data were obtained from medical records.Results: The percentage of VEGF-C positive cells in cervical diseases ranged from 0-71%, 16 (20.5%) were negative, 21(26.9%)weak positive, 26(33.4%)moderate positive andl5 ( 19.2%) strong positive. VEGF-C expression was moderate or strong positive in the 14 patients with lymph node metastases.The expression of VEGF-C and COX-2 was significantly different with the progression of cervical diseases from chronical inflammation to cervical intraepithelial neoplasia(CIN) and to invasive carcinoma of cervix uteri (p<0.05). There was a direct relationship between the expression of VEGF-C and VEGFR-3, both of them related to lymphagiogenesis and lymph node metastasis. The COX-2 positive rate in cervical carcinoma was 30.4% and a strong immunoreactive for VEGF-C and COX-2 was observed in the tumor nest. Statistical analyses suggest that the expression of VEGF-C in cervical cancer was closely related to the expression of COX-2. Moreover, the coexpression of VEGF-C and COX-2 correlated with the lymph vessel density (LVD) of the stroma adjacent to the tumor nest.Conclusions: Given these observations, we conclude that VEGF-C and COX-2 may promote the canceration of cervical cancer and that VEGF-C/ VEGFR-3 system had a significant association with the lymphagiogenesis and lymph node metastasis. COX-2 may participate in VEGF-C lymphangiogenic pathway and the high expression of them may play an important role in the lymphatic proliferation and spread in cervical cancer. Objective : To detect the expression of VEGF-C in nasopharyngeal carcinoma (NPC) and explore its relationship with proliferation and metastasis of NPC.Methods: Biopsy specimens of 62 NPC patients were divided into 2 equal portions, one for immunohistochemical staining with VEGF-C polyclonal antibody by streptavidin peroxidase method, another for flow cytometry analysis of its proliferation with Ki67 antibody. The patients were followed up periodically, and then their 3-year survival and the cause of death were statistically analyzed.Results: Of the 62 patients, the percentage of VEGF-C positive cells in the tumors ranged from 0~82% 17 (27.4%) were negative, 13 (21.0%) weak positive, 18 (29.0%) moderate positive and 14 (22.6%) strong positive. Ki67 positive tumor cells ranged from 0-52%, 40 cases (64.5%) showed LP1 which include 15 cases of negative, 22 (35.5%) showed HPI. There was a direct relationship between the expression of VEGF-C and Ki67 (r=0.323, P<0.05). The 3-year survival rale of all patients was 66.1%. The expression of VEGF-C in the patients of positive lymph node was much higher than that of negative lymph node (P <0.01). Among the 8 cases of distant metastasis patients, their VEGF-C expression was strong positive white among the 21 cases of disease free survival patients, their VEGF-C expression was (-) or (+) account for 80.9%. A inverse correlation was found between the VEGF-C expression and prognosis of NPC patients, but the difference has no statistical significant (r=-0.219, P>0.05).Conclusion : High expression of VEGF-C is related with proliferation and metastasis of NPC cells. It’s not an independent prognostic factor, but a predictive marker of disease-free survival of NPC patients.