Antitumor effects of oncolytic adenovirus expressing IL24 on bladder cancer Biu87 cells in vitro

Objective To study the antitumor effects of oncelytic adenovirus expressing human IL24 gene (ZD55-IL24) on human bladder cancer Biu87 ceils. Methods The recombinant adenoviruses ZD55-IL24 were verified by PCR. Further viruses were propagated, purified and functional PFU titens were determined by plaque assay on 293 cells. The expression of E1A and IL24 was detected by Western blot a-nalysis. The apoptosis was determined by TdT mediated dUTP nick end labeling (TUNEL). ZD55-IL24 were used to infect Biu87 cells. The inhibition of Biu87 cell proliferation by ZD55-IL24 was determined by MTI" assay and crystal violet dye method at different MOI values. Results The analysis of PCR indicated the recombinant adenovirus ZD55-IL24 containing IL24 gene without wild adenovirus. Functional PFU ti-tens of ZD55-IL24 were 2.0 × 1010 PFU/ml. Western blot analysis confirmed that ZD55-IL24 could express IL24 and E1A protein in Biu87 cell line with high efficiency. The percentage of the apoptotic cells was (52.3±3.2) %, (26.3±2.3) % and (32.0±3.1) % respectively after treatment with ZD55-IL24,ZD55-EGFP and Ad-IL24 for 48 h. Crystal violet staining and MTT assay showed ZD55-IL24 could induce obviously cytotoxic effects and growth inhibition of Biu87 cells. Cell viability rate was (27.6±1.3) %, (48.7±2.5) % and (59.7±3.53) %, respectively after tumor cells were infected with ZD55-IL24, ZD55-EGFP and Ad-IL24 at MOI = 10 on the day 4 postinfection. Conclusion The recombinant adenovi-rus ZD55-IL24 was successfully obtained. ZD55-IL24 could exert potential antitumor effects and offer a no-vel approach for gene therapy of bladder cancer. Key words: Oncolytie adenovirus;  IL-24;  Bladder carcinoma