摘要
目的观察野生型P53基因诱导人胃腺癌MGC803细胞凋亡的现象,探讨野生型P53基因导入抑制细胞增殖的机制。方法构建含目的基因P53腺病毒载体AdCMVP53,转染P53基因表达缺失的人胃腺癌细胞MGC803。应用流式细胞仪分析细胞周期和细胞凋亡率,琼脂糖凝胶电泳图谱及脱氧核苷酸末端转移酶介导的缺口末端标记(TUNEL)技术检测细胞凋亡。结果腺病毒介导的P53基因转染人胃腺癌细胞MGC803后,流式细胞仪显示757%细胞生长停滞在G0/G1期,208%的细胞发生凋亡;琼脂糖凝胶电泳可观察到呈阶梯状DNA区带图谱(DNAladder);AdCMVP53转染组可见细胞凋亡,细胞核内有特异性的蓝黑色团块。结论野生型P53基因通过诱导MGC803细胞凋亡抑制细胞增殖。
Objective To explore the inhibiting mechanism of proliferation of human gastric adenocarcinoma cell(MGC-803)by wild-type P53gene.Methods A P53gene recombinant adenovirus vector,AdCMVP53,was constructed a nd transfected into the cultured MGC-803.The cell cycle and apoptosis rate were analysed with flow cytometry.Apoptosis was determi ned by termined deoxynucleotidyl transferase-mediated d-UTP nick en d labeling(TUNEL)and agarose gel electrophoresis.Results Introduction of wild-type P53into M GC-803arrested 75.7%of AdCMVP53-infected cells at G 0 /G 1 phase of cycle,leading to inhibition of DNA s ynthesis.The apoptosis rate was 20.8%.Electrophoresis of genomic DNA showed internucleosomal fragme nt of DNA isolated from the AdCMVP53-infected MGC-803.TUNEL and flow cytometry revealed apoptosis of AdCMVP53-infected cells.Conclusion Wild-type P53gene can suppress MGC-803proliferation by i nduction of apoptosis.
出处
《中华胃肠外科杂志》
CAS
2001年第3期192-194,共3页
Chinese Journal of Gastrointestinal Surgery
