Effects of bcl-2 antisense oligodeoxynucleotide on proliferation and apoptosis of human melanoma A375 cell line

AIM: To investigate effects of bcl-2 fully phosporothioated antisense oligodeoxynucleotide(bcl-2 ASODN) on proliferation and apoptosis of human melanoma A375 cell, and its possible mechanism. METHODS: Proliferation and apoptosis of A375 cell with bcl-2 ASODN treatment were evaluated by MTT colorimetric assay, laser scanning confocal microscope (LSCM), TUNEL and Annexin V/propidium iodide(PI), and the level of bcl-2 mRNA expression in A375 cell was detected by RT-PCR before and after being treated by bcl-2 ASODN. RESULTS: MTT assay demonstrated that bcl-2 ASODN could inhibit the proliferation of the cells in both time and concentration dependent manner. Characteristic morphologic apoptosis changes were observed by LSCM after incubated with bcl-2 ASODN for 48 h. Most nucleus were labeled in brown by TUNEL in ASODN group, but not markedly labeled both in SODN and control group. The apoptosis rate of A375 cells in 30 micromol/L bcl-2 ASODN group was significantly higher than that in bcl-2 SODN and in control group. The bcl-2 ASODN-induced apoptosis of A375 cells, which was accompanied by declined expression of bcl-2 mRNA was distinctly lower than that in SODN and control groups. CONCLUSION: These results suggest that bcl-1 ASODN can not only inhibit proliferation but also induce apoptosis of human melanoma A375 cells in vitro, and the apoptosis-induced mechanism is down-regulating expression of bcl-2 mRNA.