Effect of the combined use of oncolytic adenovirus carrying tumor necrosis factor-related apoptosis-inducing ligand with epirubicin on proliferation and apoptosis of bladder cancer T24 cells

Abstract

Objective To observe the effects of the combined use of oncolytic adenovirus carrying tumor necrosis factor-related apoptosis- inducing ligand(ZD55-TRAIL) with epirubicin(EPB) on the growth and apoptosis of bladder cancer T24 cells. Methods T24 cells were divided into 5 groups: ZD55-TRAIL[multiple of infection(MOI)= 10.0] plus EPB(1 mg/L) group, ZD55- TRAIL(MOI= 10.0) group, EPB(1 mg/L)group, ZD55- enhanced green fluorescent protein(EGFP, MOI= 10.0)group, and PBS group. The expression of TRAIL was detected by reverse transcriptase- polymerase chain reaction(RT - PCR)and Western blotting. The cytotoxicity of cells was assessed by cell counting kit- 8(CCK- 8)assay. Cell apoptosis was determined by annexin V- fluoresceine isothiocyanate(FITC)assay. Results RT-PCR analysis confirmed that T24 cells treated with ZD55- TRAIL plus EPB and ZD55- TRAIL could express TRAIL mRNA with high efficiency and there was no significant difference between two groups(P> 0.05). Western blotting analysis showed that ZD55- TRAIL could effectively promote the expression of TRAIL protein in T24 cells.CCK- 8 assay revealed the survival rate after 48 h in the ZD55- TRAIL(MOI= 10.0)plus EPB(1 mg/L)group[(18.44±2.38)%] was significantly higher than in ZD55- TRAIL group[(40.75± 4.12)%], EPB group[(53.62±1.41)%]and ZD55- EGFP group[(61.65±4.53)%,P<0.01]. The single use of EPBat the 2nd day achieved the same effect as MOI= 10.0 ZD55- TRAIL+ 1 mg/L EPBat 2 mg/ L,and at 3rd day,a higher concentration of 4 mg/L was needed. The results of cell apoptosis revealed the number of apoptosis cells in ZD55- TRAIL plus EPB group was 2.01 times,2.21 times, and 2.95 times of that in ZD55- TRAIL group, EPB group and ZD55- EGFP group respectively(P<0.01). Conclusion ZD55 - TRAIL conjugated with EPB exhibited a remarkably increased apoptosis- inducing effect and great antitumor effects in bladder cancer T24 cells. Key words: Oncolytic adenovirus; Tumor necrosis factor-related apoptosis-inducing ligand; Epirubicin