摘要
目的 探讨p14ARF对喉癌细胞生长的抑制作用及其对内源性p5 3表达的影响 ,为喉癌致癌的分子机理和基因治疗提供理论基础。方法 采用基因转染技术 ,将全长野生型p14ARF 互补DNA(complementaryDNA ,cDNA)转染到人喉鳞状细胞癌Hep 2细胞中 ,利用流式细胞仪、聚合酶链反应(polymerasechainreaction ,PCR)和Western Blotting等方法研究其对细胞周期、内源性野生型p5 3表达的影响。结果 转染p14ARF基因的Hep 2细胞的生长受到明显抑制 ,其克隆形成能力为转染空载体的5 7%。细胞分析表明 ,转染p14ARFcDNA 48h后 ,处在G0 G1和G2 M期的细胞皆增加了 1倍。同时Western Blotting分析结果表明 ,内源性野生型p5 3表达明显上升。结论 p14ARF 基因能上调Hep 2细胞中内源性野生型p5 3的表达并阻滞细胞于G0 G1和G2 M期 ,从而抑制其生长。
Objective To explore the inhibitory effect of p14 ARF on the cell growth of laryngeal carcinoma and the expression on endogenous p53. Methods p14 ARF cDNA was transferred to the cell line Hep 2 of squamous cell carcinoma of the larynx by gene transfer to study the cell cycles and the expression of endogenous wild type p53 using flow cytometry, polymerase chain reaction(PCR)and Western Blotting. Results Expression of p14 ARF significantly affected the Hep 2 cell growth. The clone forming efficiency of the Hep 2 cells transferred with p14 ARF was 57%,compared with empty vector pcDNA3. The number of 48 hours after transfer with p14 ARF cDNA at both G0/G1 and G2/M was two fold as the control. The expression of endogenous wild type p53 was significantly enhanced. Conclusion Expression of p14 ARF can up regulate the expression of endogenous wild type p53 and inhibit the Hep 2 cell growth of human laryngeal squamous cell carcinoma at both G0/G1 and G2/M.
出处
《中华耳鼻咽喉科杂志》
CSCD
北大核心
2001年第2期135-137,T009,共4页
Chinese Journal of Otorhinolaryngology
