| Objective: To establish a cell line of squamous cell carcinoma(SCC) from Sprague-Dawley rat buccal mucosa induced by 4NQO. To study and identify its biological characters. The cell line provided an effective model which can be used in basal research of etiology, mechanism,diagnosis and therapy of oral carcinoma.Materials and Methods: (1)The buccal mucosa SCC models of Sprague-Dawley rats were induced by adding 4NQO into the rats' drinking water. The tumors were diagnosed and identified pathologically. In vitro tissue and cell cultures were employed to obtain mixed cells. The mixed cells were purified by using trypsinization and cell cloning , then the purified cells were passaged in vitro. (2)The biological characters of the cells were studied through inverted microscopes, scanning and transmission electon microscopes, pathological and immuno- histochemical observation, chromosome analysis, MTT technique, flow cytometry assay , and analysis of telomerase activity.(3)Xenograft inoculations of the cells(3×10~6/200μl) in nude mice provided models to observe the metastatic characters of SDSCC-B. (4)A cell subline was filtrated through continuous inoculation in nude mice, including hypodermic inoculation and injection by tail veins. Tumor growth velocity and ratios of LW/BW (lung weight/body weight) were statisticly analyzed by SAS6.12 statistical package. (5)Cells and tissues were inoculated in situ and subcutaneous incision of SD rats. Consequently, inoculation tumors were expected to be observed.Result: (1)The findings suggested that the tumors of SD rats induced by 4NQO were SCC pathologically, and the cell line was squamous cell carcinoma cells. The cells had been cultured for 11 months and 22 days, and transferred for 105 passages by March 20th, 2005. The cell line was named Sprague-Dawley rat buccal squamous cell carcinoma cell line(SDSCC-B). (2)The cells were consistent with morphology of SCC by microscopes. Cytokeratin and vimentin were both positive immunohistochemically.The population doubling time was 26 hours by using the method of MTT, and was analyzed statistically with no significant difference(P>0.05) between passage 21 and 65. The cells in S-phase occupied 17.83~22.13%, while G1-phase and G2/M-phase cells were respectively 63.34~ 70.09% and 11.77-—14.53%. The chromosome modal number was 78. Telomerase activity was... |
