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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: To investigate the relationship between the occurrence of retinoblastoma (Rb) and the deletion and inactivation of Rb gene (Rb1). Methods: The retroviral vector pDOR-Rb1 + was constructed by using retrovirus vector pDOR and full-length 4.7kb wild-type Rb1 cDNA. PDOR-Rb1 + was transfected into the CRIP packaging cell line by liposome-mediated method. Results: The experiment produced 0.5X105Cfu recombinant retrovirus with primary infection ability. SO-Rb50 was infected by the virus and screened by G418 to obtain an anti-G418 cell population. The transfected cells were detected by PCR and Southern blotting. The results showed that there was a complete exogenous Rb1 in the anti-G418 cells. Northern blot showed that the expression of Rb1mRNA increased. The determination of colony formation ability of soft agar on growth rate of cell population showed that the expression of exogenous Rb1 reduced the colony forming ability of SO-Rb50 in soft agar, but had no significant effect on population growth rate. Conclusion: Exogenous Rb1 has a certain effect on the malignant phenotype of SO-Rb50.