Shou J, Zhang Y, Wu D
Institute of Radiation Medicine, Beijing.
Chin Med Sci J. 1996 Sep;11(3):162-5.
The mutation of p53 gene was detected in Syrian hamster embryo (SHE) cells neoplastically initiated with alpha particles. The level of the p53 mRNA in transformant was obviously higher than that in non-irradiated counterpart, as measured by Northern blot analysis of total RNA. A pair of primers were designed based on p53 cDNA sequence to produce the whole length of coding sequence about 1.2 kilobase (Kb) by reverse transcription of mRNA followed by the polymerase chain reaction (RT-PCR), but the length of fragment amplified from transformant mRNA was about 0.3 Kb, remarkably shorter than that from normal SHE cells. Immunohistochemical analysis of p53 protein showed that no heavy staining was found on slice of tumor derived from transformant inoculated in nude mice with hamster specific p53 monocloned antibody HD200. The results implied that p53 gene had been mutated by deletion, which might lead to loss of p53 protein expression but the increased expression of p53 remained in alpha particle-induced SHE transformant.
在用α粒子进行肿瘤起始处理的叙利亚仓鼠胚胎（SHE）细胞中检测到了p53基因的突变。通过对总RNA进行Northern印迹分析可知，转化细胞中p53 mRNA的水平明显高于未辐照的对照细胞。根据p53 cDNA序列设计了一对引物，通过mRNA逆转录随后进行聚合酶链反应（RT-PCR）来产生约1.2千碱基（Kb）的全长编码序列，但从转化细胞mRNA扩增的片段长度约为0.3 Kb，明显短于正常SHE细胞的片段长度。用仓鼠特异性p53单克隆抗体HD200对接种于裸鼠的转化细胞来源的肿瘤切片进行免疫组织化学分析，结果显示未发现重度染色。结果表明，p53基因因缺失而发生了突变，这可能导致p53蛋白表达缺失，但在α粒子诱导的SHE转化细胞中p53的表达仍增加。
