摘要
目的探讨p53基因对胃癌细胞株BGC823中Survivin基因表达的影响及其作用机制。方法应用逆转录．聚合酶链反应(RT-PCR)和Western blot技术检测胃癌细胞株BGC823中Survivin和p53基因表达情况；利用质粒pcDNA 3．0-rp53转染该细胞株,应用实时聚合酶链反应(PCR)和Western blot检测细胞在培养后不同时间点Survivin mRNA和蛋白表达水平。结果确认了胃癌细胞株BGC823中Survivin和突变型p53基因的表达。受野生型p53质粒转染的胃癌细胞在培养16和24 h后,Survivin mRNA和蛋白质水平呈明显下降趋势。结论野生型p53在转染胃癌细胞株BCB23后可显著抑制后者Survivin基因的转录和表达。
Objective To detect the expression of Survivin and p53 genes in the gastric cancer cell strain BGC823, and to study how p53 gene influences Survivin gene expression and the presence of possible interacting biological mechanisms between Survivin and p53. Methods RT-PCR, real time quantitative PCR and Western blot techniques were applied to observe the gene expression and protein production of both Survivin and p53 in the gastric cancer Cell strain BGC823, respectively. Then the BGC823 ceils were transfected with wild-typed-p53 plasmid, pcDNA3.0-rp53, using mock-pcDNA3.0 as a control. All cells were incubated, and real-time PCR and Western blot techniques were applied to examine the levels of Survivin mRNA and protein at the intervals of 8, 16, and 24 h. Results The results of RT-PCR and Western Blot revealed that the gastric cancer strain did transcribe and express the Survivin and mutant-p53 genes. Those wild-typed-p53-processed BGC825 cells revealed no significant impact on Survivin mRNA and protein levels at 8th h after the transfection, but the mRNA and protein levels began to fall at 16th h. This tendency kept going down after 24 h of the transfection. Conclusion The wild-typed p53 gene can inhibit the transcription and expression of Survivin at 16th h after transfection, which is more apparent after 24 h of transfection.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第6期654-655,共2页
Chinese Journal of Experimental Surgery
