Abstract: Objective To establish a nude mouse model of liver cancer and explore whether drug resistance of cancer stem-like cells in vivo can lead to enrichment of stem-like cells. Methods A subcutaneous tumor model of liver carcinoma was established by subcu-taneously injecting human liver BEL-7404 cell suspensions into both axilla of BALB/C-nu mice.Following adriamycin treatment，primary cultures were established， and both parental and adriamycin-treated cells were studied in assays of single-cell colony formation and sphere cell formation to detect drug resistance and analyze other cytological characteristics. Results The tumor formation rate in this BALB/C-nu subcutaneous tumor model was 100%.There were no significant differences in the rate of single-cell colony formation between BEL-7404 parental liver cells and first-generation cells after adriamycin treatment（P>0.05）.However，second-generation cells showed a significantly higher rate of single-cell colony formation and a significantly lower colony formation rate than parental or first-generation cells（P<0.05）. Non-adherent spheres were not observed in cultures of any of the three cell types. Conclusions Low-dose adriamycin intervention may be a way to enrich for liver cancer stem-like cells in vivo.

Key words: Liver neoplasms, Cancer stem cell, Subcutaneous tumor model, Adriamycin