野生型p53基因的表达及其对子宫颈癌细胞株的生长抑制作用

The expression of wild─type p53 gene and growth supression of cultured human cervical carcinoma cells

构建了可在哺乳动物细胞中表达人ｐ５３蛋白的重组质粒ｐ５３─ｐＳＶ２ｎｅｏ，分别转染猴肾细胞株ＣＶ─１及人子宫颈癌细胞株ＳｉＨａ。用ＰＣＲ法检测出转染细胞中的ｐ５３ｃＤＮＡ；用免疫组化ＡＢＣ法检测出ｐ５３蛋白的表达定位于细胞核内，用Ｗｅｓｔｅｒｎｂｌｏｔ检测出转染细胞中存在ｐ５３蛋白。结果表明，外源性野生型ｐ５３基因的表达可以使ＳｉＨａ细胞的细胞克隆形成效减少５０％，使细胞生长速度降低，但对ＳｉＨａ细胞的软琼脂克隆形成效无明显影响。提示ｐ５３基因的肿瘤抑制作用与野生型ｐ５３蛋白的量有关，并且是细胞类型特异性的。

In this study, we construct a wild type p53 gene recombinant plasmid p53─pSV2neo, which can expresshuman p53 protein in the mammalian cells.The recombinant DNAwas used to transfect the monkey kidneycell line CV─1 and human cervical carcinoma cell line SiHa by the way of lipofectin. p53cDNA was detectedby PCR; The expression of p53 protein was detected in the nuclei of the cells by ABC technique; p53 proteincould also be detected by Western blot. In this expression we found that the SiHa cells transfected withwild type p53 gene formed colonies 50%less efficiently than those transfected with pSV2neo, and thegrowth rate was also slow.But the numbers of colonies formed in the soft agar had no significant alter─ation.This suggests that the effects of p53 may vary with both protein dosage and cell type.