BRD7基因转染对鼻咽癌细胞生长的抑制作用  

Growth Suppression of Nasopharyngeal Carcinoma Cell through Transfection of BRD7 Gene

作　　者：余鹰[1] 朱诗国[1] 张必成[1] 李忠花[1] 向娟娟[1] 周鸣[1] 李小玲[1] 李桂源[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所,湖南长沙410078

出　　处：《癌症》2001年第6期569-574,共6页Chinese Journal of Cancer

基　　金：国家863项目(No.102-10-01-05);973重点项目<"疾病基因组学"理论和技术体系的建立>(G19980510

摘　　要：目的：探讨鼻咽癌负相关基因ＢＲＤ７对鼻咽癌细胞系ＨＮＥＩ生长的影响。方法：构建ＢＲＤ７基因真核表达载体ｐｃＤＮＡ３.１（＋）／ＢＲＤ７重组体，采用脂质体介导转染技术，将ＢＲＤ７真核表达重组质粒和空载体质粒分别导入鼻咽癌细胞系ＨＮＥ１，Ｓｏｕｔｈｅｒｎ杂交和ＲＴ－ＰＣＲ分别检测外源性ＤＮＡ的整合和ＢＲＤ７基因的表达，并借助细胞生长曲线、软琼脂集落形成试验、流式细胞计数和裸鼠接种方法对转染细胞的生物学行为进行了检测。结果：转染ＢＲＤ７基因的 ＨＮＥ１生长倍增时间为５３ ｈ，较ＨＮＥ１（２３．９ｈ）和空载体转染 ＨＮＥ１（２４.１ｈ）明显延长，流式细胞仪表明，ＢＲＤ７表达升高延缓细胞由Ｇ０－Ｇ１期进人Ｓ期，ＢＲＤ７转染ＨＮＥ１在软琼脂中集落形成率较对照组显著下降（Ｐ＜０．０１），裸鼠接种试验显示ＢＲＤ７基因转染细胞ＨＮＥ１生长速度受到抑制。结论：ＢＲＤ７基因重表达有助于ＨＮＥ１的恶性表型的逆转；ＢＲＤ７是一个鼻咽癌相关的抑瘤基因良好的候选者。Objective: This study was designed to explore the effect of BRD7 gene negative-associated with nasopharyngeal carcinoma (NPC) on the growth of NPC cell line HNEI. Methods: The mammal expression vector of BRD7, pcDNA3. 1 (+ ) /BRD7, was constructed and transfected into HNE1 cell. Stable G418-resistant clones were isolated, and the integration of the exogenous vector DNA and the expression of BRD7 gene were detected by Southern blot and HT-PCR respectively. Finally the cytobiological characterization of positive clone (B-4) was analyzed by using population double time (PDT), soft agar assay, cytometry, and xenograft. Results: The PDT of G418-resistant HNE1 cell with epression of BRD7 was 53 h and significantly longer than that of vector-transfected HNE1 cell and untransfected HNE1 (P < 0. 01 ). Flow cytometric data shown that more BRD7 transfected cells went into phase G0 - G1 than controls. And it also presented decreased clonogenicity and tomorigenicity in soft agar assay and tumor formation in nude mice. Conclusion: The reexpression of BRD7 could favor the malignant phenotype revision of NPC cells. And BRD7 gene might be a good candidate of tumor suppressor gene correlated with NPC.

关 键 词：鼻咽肿瘤 BRD7 抑瘤基因 基因转染 基因表达 HNE1 NPC 

分 类 号：R739.63[医药卫生—肿瘤] R730.231[医药卫生—临床医学]