The Establishment And Characterization Of Urogenital Tumor Infinite Cell Lines

Tumor infinite cell lines are the most useful tools for the basic research of neoplasm. But the characters of the tumor cell lines will change with the long - time - culture in vitro, so it is imperative to establish new tumor cell lines. Urogenital tumor cell lines are not likely to be cultured in vitro, but the introduction of nude mice in the primary culture enhance the procedure which may be very difficult and the implanted tumor is identified to the primary tumor in morphologically, pathologically and histologically.Tumor metastasis and the immune - evade are the two distinctions of tumors and they are the main subjects of many years'research. The establishment of tumor metastasis model and the selection of high metastatic subline are the basic points for the research of metastasis. Many laboratories stated that metastasis happens seldom when tumor samples were implanted into nude mice subcutaneously, while orthotopic transplantation is "patient -like" and results clinic - mode metastasis. Further research discovered that transplanting the intacted tumor tissue orthotopically induced more metastasis than injecting tumor cell suspension into the according organs in nude mice. So the SOI (Surgical Orthotopic Implantation) model is considered to be the mosthelpful model in studying tumor metastasis.Hie studying of tumor immunology is hampered because the tumor shows low immunogeneity. But the finding of CTL which can be induced by TIL and PBL with the stimulation of autologous melanoma shed light on this field. CTL is the strongest tumor killer in the immunosystem and a series of TAA ( Tumor Associated Antigen) were discovered through CTL. Now the application of CTL and " DC - peptide" vaccine in the clinic show much effect.Using three methods; tumor tissue culture, tumor cell suspension culture , xenotransplanting tumor tissue into nude mice, we tried 43 tumor samples to establish Urogenital tumor cell lines. At last 3 infinite cell lines were successfully established by xenotransplanting tumor samples into nude mice. They are renal clear cell carcinoma RCC - 9863, bladder transitional cell cancer BC -6 and prostate tumor cell PC -98106. These cell lines are cultured in vitro for more than one year and are all identified according to the references.We then implanted the intacted RCC - 9863 tumor tissue into nude mice orthotopically. About one month, the status of nude mice decreased, while the nude mice with tumor subcutaneously lived normally. Autopsy the nude mice and found the metastasis in lung, regional lymphatic node and microme-tastasis in liver, and the primary tumor grew invasively. Cultured the meta-static tissue in lung and injected the cell suspension orthtopicaUy, a larger lung metastasis was found. Repeated this procedure and finally a high meta-static subline MRCC was selected. It express high PCNA and IV collagnase and can form lung micrometastasis even when it was injected into nude mice subcutaneously.We separated lymphocyte from PBL of the same patient from whom the RCC -9863 was established. Under the low dose IL -2 (50u/ml) , stimulated by radiated RCC -9863, the lymphocyte propagated quickly and became larger and like sommma. It has the specific cytotoxic to RCC - 9863 but no effect on GRC-1, T24, BIU-87, PC-3, LNCaP and K562. The expres-sion of CD3 and CDS are 96.7% and 64% respectively. These data proved that this cytotoxic lymphocyte was CTL. But the culture period of CTL is only 3 month and thus more research needed to modify the culture condition of CTL.Conclusion; We established three cell lines and renal cancer SOI model through nude mice. Also we selected a high metastatic subline and these may be the basis for further research of tumor metastasis. We explored a primary study of renal cancer CTL and testified that TAA exists in renal carcinoma and a thoroughly study of renal cancer peptide is optically.