重组P_(53)基因转染的肝癌细胞克隆株的建立及其生物学活性观察

人Ｐ５３基因位于１７ 号染色体短臂上（１７ｐ１３．１） ，编码一个由３９３ 个氨基酸组成的核磷酸蛋白，有调控细胞的生长，维持基因组的稳定，其突变、缺失在恶性肿瘤发生发展中起着非常重要的作用。为进一步探讨野生型Ｐ５３ 基因的抗肿瘤特性，我室采用分子克隆方法构建了重组野生型Ｐ５３ 真核表达载体（ｐｘＴ１—Ｐ５３） ，用磷酸钙—ＤＮＡ 共沉淀法将其导入人肝癌ＨｅｐＧ ２ 细胞内，通过标记基因ＮｅＯＲ 筛选带外源Ｐ５３ 基因的Ｇ４１８ 抗药性克隆，对Ｇ４１８ 抗药性克隆细胞的生长增殖和粘附能力进行观察。结果表明，导入野生型Ｐ５３基因能使肝癌ＨｅｐＧ ２ 细胞的生长增殖速度较对照细胞明显减慢，细胞粘附能力下降。提示野生型Ｐ５３ 基因能抑制肝癌细胞的生长。本研究为肝癌的Ｐ５３ 实验性基因治疗提供了一定的依据。

Observation of the Biological Characteristics of the Tumor Cells Transfected with Recombinant P 53

P 53 gene is located on the short arm of human chromosome 17 at position 17p13.1. This gene encodes a 393 amino acid nuclear phosphoprotein which involves in the regulation of cell proliferation. Loss of normal P 53 function is associated with the cell transformation in vitro and the development of neoplasms in vivo. More than one half of human malignancies were shown to contain an altered P 53 gene. To gain more insight into the functional role of wild type P 53 in human carcinoma, we constructed recombinant wild type pxT 1 P 53 expression vector. We performed experiments to transfer wild type P 53 gene into human hepatoma cell line(HepG 2) with the calcium phosphate precipitation method. We assessed cell growth properties and the changes of adhesive. The results demonstrated that human wild type P 53 gene can suppress the phenotype of HepG 2 cell line. These results encourage us to do further experiments on tumor cell targeted P 53 therapy.