Objective: To isolate and characterize cancer cell subclones with different metastatic potential from human metastatic prostate cancer cell line (PC-3M).
Methods: Using limited dilution, in vitro growth, Matrigel invasion assay, soft agar cloning and in vivo tumorigenicity and spontaneous metastasis assay in nude mice, were isolated and characterized four subclones with different metastatic potential.
Results: Four subclones derived from PC-3M were 1E8, 2E7, 2B6, 2B4. Each subclone exhibited a different metastatic potential when inoculated into nude mice. Among these subclones, 1E8 expressed as the highly metastatic phenotype, with 100% metastasis frequency 5 weeks after subcutaneous inoculation into nude mice, whereas 2B4 was not metastatic. In vitro 1E8 was found to be the most highly invasive cell line in Matrigel invasive assay (98 +/- 24) and had the most clones in soft agar cloning assay (265 +/- 39) while 2B4 was found to have the least invasive abilities (12 +/- 4) and the least clones (137 +/- 14).
Conclusion: Successful establishment of these subclones with different metastatic potential may be valuable for further study on the molecular mechanisms of cancer metastasis and cloning of cancer metastasis-related genes.