Fig. 6.
PCDH-PC promotes anchorage and androgen-independent CaP cell growth, a: Soft agar assays of PC-3 stably expressing PCDH-PC (PC-3/PCDH-PC) or mock transfected PC-3 (control PC-3) cells. Cells (20,000) were seeded in triplicate in 6-wells plates. The colonies were counted after 2 weeks of culture in completed medium (RMPI/5% FCS). The number of soft agar colonies presented is the mean of colony counts in 10 40× microscopic fields from three wells, b: Growth curve assays. LNCaP/PCDH-PC cells or empty-vector-transfected LNCaP cells (3,500) were seeded in triplicate in 12-well plates. The cells were cultivated in hormone-deprived medium (RMPI/5% CS-FCS). At 2–3 day intervals, the cell number was analyzed. In contrast to control cells, LNCaP/PCDH-PC continued to proliferate in androgen-depleted medium, c: Anchorage-independent growth in hormone-deprived medium. LNCaP/PCDH-PC cells or empty-vector-transformed LNCaP cells (20,000) were seeded in triplicate in 6-well plates. The colonies were counted after 2 weeks of culture in RMPI/5% CS-FCS. Error bars indicate standard errors. *P < 0.001 compared with control-cells. d: Tumorigenicity of PCDH-PC overexpressed LNCaP cells in castrated nude mice. 2 × 106 of either control LNCaP cells or LNCaP/PCDH-PC cells were injected into eight nude mice castrated 1 week before injection. After 7 weeks, mice injected with control cells had no visible or palpable tumor (0/8) whereas 100% of castrated mice xenografted with LNCaP/PCDH-PC cells formed tumors (8/8). Hematoxylin and eosin staining showed these tumors were highly vascularized. Magnification: 200×.