Experiment Studies On The Establishment Of The Bear-tumor Mice Model And Gene Array Differential Expression Between Local Tumor In Situ And Metastatic Tumor Of Neuroblastoma

Objective: By establishing the bear-tumor nude mice model of transplanted tumor in situ and metastatic tumor, heterogenicity of tumor theory is proved, and so, we can discuss the metastasis mechanism of neuroblastuma deeply.Methods: Took the fresh tumor tissue from the patient under the sterilizing: situation during the operation, used the enzyme digestion method to establish the in vitro cell line. We inoculated the single cell suspension by the mode of hypodermic injection for 11 SPF nude mice and 3 SCID mice, established the bear-tumor nude mice model of local tumor in situ and metastatic tumor, then use the in vitro cell line of the bear-tumer nude mice model of local tumor in situ and the human in vitro cell line we had established to inoculate the single cell suspension by the mode of hypodermic injection for 14 nude mice and the mode of celiac injection for 6 nude mice. The injected dose is 0.5 ml in the consistence of 2.5×10~7/ml.Results: We obtained the bear-cancer nude mice models (5/14) and the SCID mice models (2/3). There are two nude mice with metastasis all over the body, the others obtain the tumor at inoculated site. The second time all the nude mice totally bear tumor. In the human in vitro cell line hypodermic injection group, there are 5 nude mice wita metastasis, the others obtain the tumor at inoculated site; In the mouse in vitro cell line hypodermic injection group, all the 6 mice bear the tumor at inoculated site; in the human in vitro cell line celiac injection group , all the 6 nude mice are with metastasis in abdominal viscera. All were testified by pathology.Conclusions: we successfully establish the bear-tumor mice model of local tumor in situ and metastatic tumor twice. It proved that our human in vitro cell line we had established has local oncogenesis and metastasis character, and metastasis is organ specific. So heterogenicity of tumor theory is proved. Our experiments can supply a very important basis for the further research to the metastasis mechanism of neuroblastuma. Objective: By comparing gene expressions of the local tumor in situ with metastatic tumor of bear-tumor nude mice model, search the related metastatic gene.Methods: 4 SuperArray series Oligo Tumor Metastasis Microarray were divided 3 group, hybridize the local tumor in situ and metastatic tumor of bear-tumor nude mice model, detect photogenesis signal, by analyzing image and data ,find the differential gene between the local tumor in situ and metastatic tumor.Results: In 113 candidate gene, we screen 25 signifcant different expression genes between local tumor in situ and metastatic tumor of neuroblastoma, including Cell adhesion genes ;Apoptosis genes ;Cell cycle genes; Matrix metalloproteinases; Cell growth and proliferation genes; Apoptosis genes.Conclusions: It exists significant difference in gene expression between local turnoi in situ and metastatic tumor of neuroblastoma that were established by the same cell line. these genes maybe closely correlate with neuroblastoma metastasis. These genes may be characteristic matastasis genes of neuroblatoma, so it's worth to advanced study.