Fig 2.

Tumorgenicity properties (A) After incubating in urine for 1 hour, 1x10(6) living cells were collected for proliferation, adhesion, invasion and colonies formation assay. CaB and non-CaB urines showed similar toxicity in all four cell lines. Data was presented as relative value against culture medium control. (B) CaB U41 and non-CaB U63 were showing significant result on all four cell lines. For proliferation assay, living cells were evaluated by MTT. U41 promoted cell proliferation and U63 suppressed or killed all cells for all four cell lines. (C) For adhesion assay, adhesion ability was evaluated by measuring the attached cells by MTT assays and OD reading was proportional to the number of adhered cells. U41 preserved the adhesion ability of HTB9, RT4 and T24, while U63 inhibited the cell adhesion of RT4 significantly. (D) To evaluate the invasion ability, cells passing through the matrigel were stained with Crystal Violet and total number of invasive cells were counted using the ImageJ software. (E) Colonies formation abilities were determined by seeding 1000 living cells onto a 100mm plate and incubated for 9 days. Colonies were then stained with crystal violet and colonies with more than 50 cells were counted. All data above was presented as relative value against culture medium control and all assays were done in triplicate.