Establishment And Study Of Retransplantation Human Oral Verrucous Carcinoma In Nude Mice And With Its Expression Of Skp2 And P27

OBJECTIVE: To establish a xenografted model of oral verrucous carcinoma in nude mice, then find the best method to establish the model, and the last aim is provide an ideal model for clinic therapy and basic research of oral verrucous carcinoma.METHOD: A fresh human oral verrucous carcinom specimen was obtained from a patient under aseptic condition during operation,included primary and metastatic tissue. Part one:Using parts of primary tissue, quickly cutting these into 1×1×1mm~3 pieces, cultured in hanks medium, and subcutaneously transplanted into Balb/c- nu/nu mice. Using some other parts of primary tissue, make it as suspension cell line, then subcutaneously injected into the right flank of the test mouse.Part two: Using parts of metastatic tissue, quickly cutting these into 2×2×2mm~3 pieces, cultured in hanks medium, and subcutaneously transplanted into Balb/c- nu/nu mice. The mice were observed every 5 days, and the growth of tumor was recorded and growth curves were figured, were sacrificed when they showed serious illness, the xenograft tumors were finally removed for histopathological study and the mice were laparotomized to examine metastatic tumors in liver and lungs and so on.RESULTS: Part one: The position where tissue block is transplanted subcutaneously displays red swelling, but the mass there is not enlarged; then, the transplanted tissue is absorbed gradually and disappears at last. The tumors in the nude mice which are injected with suspension do not grow either, with only the subcutaneous trace of injection left.Part two: Seven nude mice have growing masses where tissue blocks are imbedded subcutaneously. The tumor formation rate is 87.5%, since there are altogether eight mice. The appearance of transplanted tumors: quasi-circular; those tumors in larger volume show telangiectasia, ulcer and incrustation in their surfaces. They are movable and adherent slightly to the inner surface of the nude mice's skin, so they can be easily separated from the skin. They have smooth surfaces and complete capsule. The cut section had hard texture and a fish-flash cut surface, and some individual necrotic ones have tofu-pulp cut surface. HE staining shows that the cancer cells of those model tumors and mesenchymal components remain morphologically tumor-induced. The liver, renal, lung and lymph node do not show obvious metastasis. The immunohistochemical examined of S- phase kinase associated protein 2(Skp-2) and kinase inhibited protein(p27) was same as it in tissue of person.CONCLUSION: 1. The second part of oral verrucous carcinoma and squamous cell carcinoma metastasis model is successful, and with a higher survival rate, the model is the same of the human tumor; 2. Use fresh carcinoma tissue in nude mice through the cultivation of the model is feasible.