Acceleration of apoptosis by transfection of Apoptin gene in retinoblastoma cells / 眼科研究

ABSTRACT

Objective Present study aimed to observe the effects of Apoptin gene on killing retinoblastoma HXO-RB_(44) cells and illustrates its mechanisms. Methods Human retinoblastoma cells strain, HXO-RB_(44), was cultured and passaged in RPMI 1640 medium containing bovine serum. Apoptin gene was transfected into HXO-RB_(44) cells by liposome into HXO-RB_(44)/Apoptin, and pcDNA_3 was transfected in HXO-RB_(44)/peDNA_3 group. The expression of Apoptin mRNA was detected using Reverse Transcription Polymerase Chain Reaction (RT-PCR). The expression of protein of Apoptin and p53 were detected by SABC immunohistochemistry. The growth rate of HXO-RB_(44) cells was studied by constructing the growth curve and calculated as the formula inhibitory rate = 1-cell number in experiment group/cell number in control group x 100%. Cellular apoptosis was determined by flow cytometry. Results The RT-PCR result showed the 450 kb specific band in UXO-RB_(44)/Apoptin group and absent amplification result in HXO-RB_(44) group and HXO-RB_(44)/pcDNA_3 group. The difference in SABC-positive cell number between HXO-RB_(44)/Apoptin group and control group was statistically significant (P ＜ 0. 05). The growth of HXO-RB_(44) cells was significantly inhibited in HXO-RB_(44)/Apoptin group compared with control group (P ＜ 0.05). Apoptosis cells increased significantly. The apoptosis rate was 38. 5% . Conclusion Apoptin gene could inhibit the growth of HXO-RB_(44) cells effectively. Up-regulation of expression of p53 gene might not be one of cell apoptosis mechanisms.