Rb基因对人视网膜母细胞瘤株SO-Rb50的影响  

EFFECTS OF Rb_1 ON THE GROWTH OF HUMAN RETINOBLASTOMA CELL LINE SO-Rb50

作　　者：徐斌[1] 王新娟[1] 高汉林[1] 吕向东[2] 廖晓蓓 易玉珍[4] 

机构地区：[1]北京医科大学生化与分子生物学系,100083 [2]中国医学科学院基础所 [3]美国NIH [4]中山眼科中心

出　　处：《北京医科大学学报》1997年第5期425-428,共4页Journal of Peking University(Health Sciences)

基　　金：国家自然科学基金!39270761

摘　　要：目的：探讨视网膜母细胞瘤（Rb）的发生与Rb基因（Rb1）缺失、失活等异常的关系。方法：用逆转录病毒载体pDOR与全长4．7kb野生型Rb1cDNA构建逆转录病毒表达载体pDOR－Rb1+。用脂质体介导法将pDOR－Rb1+转入CRIP包装细胞系。结果：实验产生了0．5X105Cfu具有一次感染能力的重组逆转病毒。利用该病毒感染SO-Rb50，经G418筛选，获得了抗G418细胞群体。运用PCR及Southern杂交技术对转染细胞进行检测，结果表明该抗G418细胞中有完整的外源Rb1存在。Northern杂交发现其Rb1mRNA表达水平有所提高。对细胞群体生长速率、软琼脂集落形成能力的测定表明，外源Rb1的表达使SO-Rb50在软琼脂中集落形成能力降低，而群体生长速率无明显影响。结论：外源Rb1对SO-Rb50恶性表型有一定影响。Objective: It is known that Rb1 plays an important role in the development of retinoblastoma(Rb).A further study is supposed to investigate the close relationship between Rb1 and Rb. Methods: A retro-virus vector containing wild-type Rb1 has been constructed. The recombined vector was introduced by lipofectin into amphotropic package cell line CRIP and 0. 5 x 105 cfu retroviruses were produced. After supernatants containing the virus infected SO-Rb50, PCR, Southern and Northern blot were used to testify the existence of exogenous Rb1, soft agar and growth rate assay were used to study the effects of Rb1 on malignant phenotype of the reconstituted cells. Results: PCR and Southern blot showed that exogenous Rb1 existed in transfected cells. Northern blot showed that the level of Rb1 mRNA increased in transfected SO-Rb50. The ability of the cells to form colony in soft agar was reduced. However, no changes in growth rate were observed.Conclusion: The malignant phenotype is affected by the introduction of exogenous Rb1.

关 键 词：视网膜母细胞瘤 病因学 基因转染 RB基因 

分 类 号：R739.7[医药卫生—肿瘤]