脂质体介导p27基因转染对喉癌细胞系Hep-2生长影响的研究  

Effects of exogenous p27 gene on the cell cycle and proliferation of Hep-2 laryngeal cancer cell line

作　　者：孙安[1] 孙永柱[2] 高卓平[1] 席爱民[1] 段学军[1] 陈谊辉[1] 

机构地区：[1]西安市第四医院耳鼻咽喉科,陕西西安710004 [2]第四军医大学唐都医院耳鼻咽喉科,陕西西安710038

出　　处：《中国耳鼻咽喉头颈外科》2009年第11期632-635,共4页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：陕西省自然科学基金资助项目(SJ08-ZT09)

摘　　要：目的探讨p27基因对喉癌Hep-2细胞系生长的抑制作用,为喉癌发生机制的研究和基因治疗提供理论依据。方法采用基因转染技术,将p27cDNA转染到喉癌Hep-2细胞系中,了解其对细胞增殖能力及细胞周期的影响。结果转染p27基因的喉癌细胞生长受到抑制,其在软琼脂上克隆形成能力降低了大约4倍。对细胞周期分析表明,处在G0～G1期的细胞由32.2%增加到66.9%,经Dotblot、Western blot杂交和免疫组化证实,p27mRNA表达有明显差异(P<0.01),p27的蛋白表达量有明显差异(P<0.01),说明p27蛋白具有生长抑制功能,其作用点为G1～S期。结论提高喉癌细胞中p27的表达,能抑制肿瘤细胞的生长,导入p27基因是一种治疗喉癌的新途径。OBJECTIVE To explore the effect of p27 gene on the growth inhibition of laryngeal carcinoma cell line Hep-2. METHODS The p27 cDNA was transfected into human laryngeal carcinoma cell line（Hep-2 cells）with lipofectamine. The growth rate, cell cycles and colony formation rate of transfected cells were observed by means of MTT and FCM assay. RESULTS Expression of p27 in Hep-2 was identified by Dot blot, immunohistochemical method and Western blot analysis. The growth rate of Hep-2 transfected with p27 gene was markedly suppressed. Colony formation in soft agar was also decreased significantly. Cell cycle analysis by flow cytometry showed that the number of cells in G0～G1 phase of Hep-2 cells was significantly increased, while cells in S and G2＋M phase was decreased compared with that of the control Hep-2 cells. CONCLUSION Transfection of p27 gene into lower expression cancer cells can restore its suppressive effect on cell growth by arrest of cell cycle at G1 phase.

关 键 词：喉肿瘤 基因 肿瘤抑制 基因表达 转染 

分 类 号：R739.65[医药卫生—肿瘤] R735.34[医药卫生—临床医学]