Isolation and identification of side populations from lung metastatic hepa-tocellular carcinoma in rats
AIM: To establish the methods of isolating and identifying the stem-cell-like hepatocytes from rat pulmonary metastatic hepatocellular carcinoma. METHODS: Rat model of hepatocellular carcinoma with pulmonary metastasis was established by diethylinitrosamine. Hepatocellular carcinoma cells from pulmonary metastasis were proliferated by tissue culture. Side population (SP) cells and non-side population cells were isolated from the cultured cells by Hoechst33342 staining with FACS and were followed by identification for the expression of tumor stem cell markers c-kit and Sca-1 with FACS. Soft agar clone formation assay was performed to determine the tumorigenesis capacity of SP and non-SP cells from hepatocarcinoma cells. RESULTS: The rat model of hepatocellular carcinoma with pulmonary metastasis was established and stable hepatocellular carcinoma cell line was harvested from the model. Side population cells accounted for 0.5% of the total pulmonary metastatic cells. c-kit and Sca-1 were highly expressed on SP cells, with the expression rate of 85% and 75% respectively in SP cells, and 8% and 3% respectively in non-SP cells. Soft agar clone formation rates were 69.90% and 15.25% respectively in SP and non-SP cells, showing that the tumorigenesis capacity of SP cells was higher than that of non-SP cells (P ＜ 0.01). CONCLUSION: SP pulmonary metastasis hepatocellular carcinoma cells display the features of tumor stem cells.
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NSTL
万方数据
维普
