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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 observe the apoptosis of human gastric adenocarcinoma MGC803 cells induced by wild-type P53 gene and to investigate the mechanism of wild-type P53 gene introduction inhibiting cell proliferation. Methods The human adenocarcinoma cell line MGC803 was transfected with the p53 adenovirus vector containing AdCMVP53. Flow cytometry was used to analyze the cell cycle and apoptosis rate. Agarose gel electrophoresis and deoxynucleotide transferase-mediated nick end labeling (TUNEL) were used to detect apoptosis. Results After adenovirus-mediated P53 gene transfection into human gastric adenocarcinoma cell line MGC803, flow cytometry showed that 757% of the cells were arrested at G0/G1 phase, and 208% of cells had apoptosis; agarose gel electrophoresis was observed. Ladder DNA map (DNA ladder); AdCMVP53 transfection group showed apoptosis, specific nuclear blue-black mass in the nucleus. Conclusion The wild-type P53 gene inhibits cell proliferation by inducing apoptosis of MGC803 cells.