Objective：To set up a higher tumorigenicity cell line and investigate its biological characters. Methods: A prostate cancer cell line LNCap cell was co-transfected with lentivirus vector PGK-luciferase-GFP, then repeated subcutaneous injection of the cell with Matrigel followed by isolation of cells from tumors has yielded a cell line， sub-clone cell. Use CCK8, transwell migration assay to detect differences between the parental cell line and sub-clone cell. LNcap cell and sub-clone were injected subcutaneously. Tumor in subcutaneously was evaluated by bioluminescent imaging, histological and immunohistochemical staining analysis. Result: Comparing to the parental cell line, the sub-clone cell line grew vigorously and rapidly , up-regulated the ability of migration and higher tumor formation rate；Higher hot spot of bioluminescence was detected by vivo imaging，nuclear division can be see by microscopically, and also AR staining is positive. Conclusions: The sub-clone cell has more malignancy than LNCap.
