The Effects Of VEGF Monoclonal Antibody On The Growth Of Human Tongue Squamous Cell Carcinoma Cell Tca-8113 And Transplanted Tumor

Tongue squamous cell carcinoma (TSCC) is the most common malignant tumors in oral & maxillofacial region and presents severe malignancy, rapid growth and extensive invasive ability. The carcinoma can easily infiltrates tongue muscles,limits tongue movement and therefore interfere oral functions such as eating, swallowing and speaking[1]. Traditional treatment methods of this disease include surgical resection, radiotherapy and chemotherapy. Although these methods are used in clinic for treatment of most cases, other treatment methods are still under investigation to get better results.More and more evidences show that among many impacting factors, angiogenesis is key factor influencing tumor metastases and growth. As the most impotent stimulus of angiogenesis, vascular endothelial growth factor(VEGF)plays a crucial role in growth of malignant tumor. Therefore, many scholars tried to treat the malignant tumor with VEGF monoclonal antibody and some preferring results had contained. However, the inhibiting effect of VEGF monoclonal antibody on malignant tumors in oral & maxillofacial region, especially on tongue squamous cell carcinoma, is far under investigated and much more studies should be performed to elucidate its function and to explore the molecular mechanism of its inhibiting effect.Base on above knowledge, in vitro and in vivo experiments were conducted in this study to explore the effect of VEGF monoclonal antibody on growth of TSCC cell Tca-8113 and its mechanism on tumor inhibition. The results of this study will promote clinical use of VEGF monoclonal antibody and provide new clues for gene therapy and pharmaceutical treatment of this disease.Objective: To investigate the effect of VEGF monoclonal antibody on growth of TSCC cell Tca-8113 in vitro and growth of transplanted tumor in vivo, and to explore its mechanism on tumor inhibition.Methods: TSCC cell Tca-8113 was cultured in vitro and the cells were divided into control and experimental groups. The cells in experimental group were treated with VEGF monoclonal antibody while the cells in control received placebo. At different time points, cell proliferation rate, cell cycle and expression of VEGF were detected by MTT test, Flow cytometry and Real-time PCR, respectively, to verify the inhibition effect of VEGF monoclonal antibody on Tca-8113 growth and the mechanism of the effect. At same time, Tca-8113 cells were transplanted into back of nude mice to establish transplanted tumor model (N=10). The animals were divided into two groups (n=5): experimental animals received injection of VEGF monoclonal antibody in and around transplanted tumors and control animals were injected with physiological saline in 28 days.Size and weight of transplanted tumor, and animals` body weight were examined at different time points. Immunohistochemical examination was also performed to detect expression of VEGF in tumors. The purpose of the above in vivo experiment was to verify the inhibition effect of VEGF monoclonal antibody on tumor growth and to elucidate that VEGF monoclonal antibody exerts its effect by down-regulating VEGF expression and decrease angiogenesis in tumor.Results:(1) MTT assay showed that cell proliferation in experimental group was decreased significantly when compared to control cells (p<0.01). (2) Flow cytometry exhibited that cell cycle were significantly changed after VEGF monoclonal antibody treatment and G2 phase in experimental cells (7.393 ±0.323) was significantly higher than that in control cells (2.733±0.055) (p<0.01). (3) Real-Time PCR detection indicated that mRNA level of VEGF in experimental cells were significantly decreased when compared to control cells (p<0.01). (4) In vivo experiments indicated that both tumor size(p<0.01) and tumor weight(p<0.05) were significantly lower in experimental animals than their controls, VEGF staining intensity were also significantly increased in experimental animals(p<0.05).Conclusion:1. VEGF monoclonal antibody can significantly decrease proliferation of TSCC cell Tca-8113 in vitro, change cell cycle and down-regulate VEGF expression.2. VEGF monoclonal antibody can significantly inhibit growth of transplanted tumor and decrease VEGF expression in vivo.3. VEGF monoclonal antibody may exert its effect by directly inhibition proliferation of tumor cells and decrease angiogenesis in tumor.