| Background:The sarcoma is one of the primary highly malignant tumors, it origins in the mesenchymic (including contacts organization and muscle)。Occurs in the skin, hypodermic, periosteum and both sides of long bone。The sarcoma grows rapidly, in the late period tumor common necrosis, the hemorrphage, the cut face is puttying red, the nature isthin just like living fish flesh shape, the hematogenous metastasis can happen in the early time Osteosarcoma is the most common bone malignant tumor, it takes2.4%of malignant tumors in the children, the peak period of osteosarcoma occurs is10years to20years, the predilection position of osteosarcoma is the active growth epiphysis end, as distal femur and proximal tibia.As the character of creation of fusiform matrix cell, osteosarcoma grow rapidly, the course of an illness is short, at the begging it grows in the cortex, may gradually to the medullary cavity of bones, sometimes outward break the periosteum. invade into the mesenchymal tissue of surrounding soft tissue, and pulmonary metastasis always occurs before the detection of illness in the clinical.80%of osteosarcoma were recognized as pulmonary metastasis exist. But the detection ratio is8-15%nowadays. Most osteosarcoma patients have highly mutilation and prognosis mala. The neoadjuvant chemotherapy had same break these years, but the prognosis is still unfavorable, the therapy of osteosarcoma remains a complict question.Objective:First:To establish an RFP-tagged143B osteosarcoma orthotopic nude mice model of human osteosarcoma growth and spontaneous pulmonary metastasis, and could be used for monitor the growth of osteosarcoma and evaluate the antitumor drug; Second:Study the rh-endostin in the RFP-tagged143B osteosarcoma orthotopic nude mice model.Method:First:human osteosarcoma cells of143B were transfected with incasing cells PT67contained pLNCX-DsRed-2, RFP-tagged osteosarcoma cells were injected into the proximal tibia of athymic mice, the growth and to obtain of osteosarcoma was observed and analysed with whole body optical imaging system。 Second:Study the rh-endostin in the RFP-tagged143B osteosarcoma orthotopic nude mice model, Validate the effectof rh-endostin in the orthotopic osteosarcoma nude mice model, it contains two part. Part1:Establish15RFP-tagged143B osteosarcoma orthotopic nude mice model of human osteosarcoma,15nude mice were random divided to3groups after80%of the nude mice tumor growing observing by thewhole body optical imaging system. Five nude mice each group:negative control group treated with normal saline, low dose rh-endostin (0.75mg/kg) group, high dose rh-endostin (1.5mg/kg) group. The administration mode of three groups was intraperitoneal injection. In vivo imaging was taken twice per week, observing the change of fluorophore, traditional method measure gross tumor volume, compare restrain ratio of tumor between the3group. Part2:Establish20RFP-tagged143B osteosarcoma orthotopic nude mice model of human osteosarcoma,20nude mice were random divided to4groups after80%of the nude mice tumor growing observing by the whole body optical imaging system. Five nude mice each group: negative control group treated with normal saline, single rh-endostin (0.75mg/kg) group, single cisplatin (5mg/kg) group, rh-endostin combined cisplatin group. The administration mode of three groups was intraperitoneal injection. In vivo imaging was taken twice per week, observing the change of fluorophore by the whole body optical imaging system, traditional method measure gross tumor volume, stop the treatment at3weeks after the administration, the observation was taken until the nature died of the nude mice with osteosarcoma, the statistics of survival time in the different intervention study was taken.Result:The stable red fluorescent protein (RFP) expressing143B cells were established and the cell line was used to duplicate the implanted tumor model, the dynamic tumor growth process could be observed by whole body optical imaging system, and the effect of treatment to the tumor could be observed too.After3weeks treatment in part1, The high dose rh-endostin (1.5mg/kg) group has no significant difference compared with control group(P>0.05); The low dose rh-endostin (0.75mg/kg) group has significant difference compared with control group(P<0.05). The high dose rh-endostin (1.5mg/kg) group has no significant difference compared with low dose rh-endostin (0.75mg/kg) group(P>0.05); The ratio of tumor metastasis in low dose rh-endostin (0.75mg/kg) group and high dose rh-endostin (1.5mg/kg) group has no significant difference(P>0.05); In the survival time test, the mean survival time of rh-endostin group is30days longer than control group, it suggest that the rh-endostin may improve the body condition of the tumor nude mice; there suggest no significant side effect and toxicity in all the rh-endostin treatment group and control group through the detection of the body weight and body condition of tumor nude mice; and the all of the5mice of cisplatin group idied in1week after the treatment of cisplatin, it suggest that the dose5mg/kg of cisplatin has significant toxicity to the nude mice。Conclusion:This clinically relevant model of human osteosarcoma provides varying degrees of tumor growth at the primary site and metastatic potential.lt could continuous detection by whole body optical imaging system, Thus, this orthotopic model should be a valuable tool to investigate factors that promote or inhibit osteosarcoma growth and/or metastasis.The method of this osteosarcoma model is reliable, simple, replicate。The single use or combined therapy of cisplatin has significant toxicity to the143B osteosarcoma, the single use or combined therapy of rh-endostin has no significant effect to suppress the pulmonary metastasis of143B osteosarcoma, but it significant prolong the survival time of143B osteosarcoma, but the mechanism was far beyond research。... |
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