Establishment Of An Orthotopic Model Of Human Tongue Squamous Cell Carcinoma In Nude Mice And The Study Of Anticancer Drug Sensitivity

Object:1. To establish an orthotopic model of human tongue squamous cell carcinoma in nude mouse, simulated human body environment, observed the tumous cell biological behavior and condition of lymph node metastasis in vivo.2. Screening anticancer drug by MTT assay to explore the difference between human tongue carcinoma cell and nude mouse primary cell.Mothod:1. The tca83cell were injected into the left margin of tongue in nude mouse. Every location was injected with105/0.05ml. Check and record the weight, the condition of tumour growth, the length and width of tumour every two day. Mice was sacrificed when they died or their weight were less than25%of primay data. The tongue, lymph nodes were stained with hematoxylin and eosin for microscopic observation, and evaluated for histopathology.2. Tca83cell suspension was collected into sterile96-well flat-bottomed microtiter plates with the concentration of1×10-5/ml (200μl/well). Fresh tumor tissue obtained from the surgically reseceted specimens in nude mouse. the tumour was cut into pieces and pass through NO.200stainless steel meshes respectively into a complete medium. Cell suspension was collected into sterile96-well flat-bottomed microtiter plates (1×105/well) added chemotherapeutic agents PYM, Paclitaxel, Epirubicin or Cisplatin20μl per well and interacted48h. The drug concerations used were50μ·g/ml、10μg/ml、5μg/ml、1μg/ml、100ng/ml、25ng/ml、10ng/ml、1ng/ml、0.1ng/ml. After incubation, MTT solution was added to each well at a final concertration of1mg/ml pre well and the palates were incubated in another4h. DMSO was added to each well to sisslove the formazan and absorbance was read at570nm using a spectrophotometric microplate reader. Count the inhibition rate of cell growth. Result:1.Tumour was formed in6th-13th day after injected tumour cell. Tumor formation rate was92.3%, The result of HE and Immunohistochemical showed cervical metastatic rate was39.47%。2. The sensitivity of drug between the two cell are different,but there was no significant different in order of drug sensibility. The order was Paclitaxel> Cisplatin> Epirubicin>PYM.Conclusion:1. The establishment of a orthotopic mode human tongue squamous cell carcinoma in nude mouse is successful. It supplied an important experiment medium in studying tongue cancer development. Although injected Tca83cell in nude mouse didn’t have a ideal rate of lymph node metastasis, the result is more close to clinic characters and imitated the process of development of human head and neck carcinoma. The study provided useful information in establishing tongue cancer with high rate of lymph node metastasis.2. Drug sensitivity assay like a guider in clinical chemotherapy, the MTT assay showed tongue cancer cell strain and tumour cell in model had not changed in biological behavior and sensibility of drug. Such model was good study model in simulating human tumor development. It provided important information for the study of resistance genes and anticancer drugs.