CAJ | 학술논문

目的观察Apoptin基因对人视网膜母细胞瘤细胞株HXO-RB_(44)的促凋亡作用,并探讨其可能的机制.方法用脂质体将Apoptin基因导入HXO-RB_(44)细胞,通过RT-PCR法检测Apoptin mRNA的表达,同时用SABC免疫组织化学法分析Apoptin和p53的表达.采用细胞计数法检测细胞生长抑制率,流式细胞仪检测细胞周期的变化.结果转入Apoptin基因后,HXO-RB_(44)细胞的生长明显受抑制(P＜0.05).细胞周期分析可见凋亡峰,凋亡率为38.5%.细胞中可见Apoptin阳性表达(P＜0.05),p53表达差异无统计学意义(P＞0.05).凋亡细胞在荧光显微镜下形成凋亡小体.结论转入Apoptin基因可显著促进HXO-RB_(44)细胞的凋亡,Apoptin诱导的凋亡不依赖功能性p53的生成.
목적 관찰Apoptin기인대인시망막모세포류세포주HXO-RB_(44)적촉조망작용,병탐토기가능적궤제.방법 용지질체장Apoptin기인도입HXO-RB_(44)세포,통과RT-PCR법검측Apoptin mRNA적표체,동시용SABC면역조직화학법분석Apoptin화p53적표체.채용세포계수법검측세포생장억제솔,류식세포의검측세포주기적변화.결과 전입Apoptin기인후,HXO-RB_(44)세포적생장명현수억제(P＜0.05).세포주기분석가견조망봉,조망솔위38.5%.세포중가견Apoptin양성표체(P＜0.05),p53표체차이무통계학의의(P＞0.05).조망세포재형광현미경하형성조망소체.결론 전입Apoptin기인가현저촉진HXO-RB_(44)세포적조망,Apoptin유도적조망불의뢰공능성p53적생성.
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.