大肠癌HT-29,Lovo细胞p53转录表达差异同步研究

Synchronous studies on variations of p53 gene transcriptions and expressions in colorectal carcinomas HT-29 and Lovo cell lines

目的探讨HT-29,Lovo两种大肠癌细胞中p53基因mRNA转录及蛋白表达水平与其分子生物学特性之间的关系。方法以RNA真空转移法行Northern blot分析,以光敏生物素标记的探针行细胞原位杂交,对此两种细胞中p53 mRNA转录水平分别进行比较研究。采用免疫细胞化学染色方法同步对此两种细胞中P53蛋白表达进行比较研究。结果 Northern blot与原位杂交检测结果一致。两种细胞中均为p53 mRNA转录阳性,前者强度明显高于后者。P53蛋白仅在HT-29细胞检出,Lovo细胞为阴性。结论两种细胞内存在不同类型p53基因,HT-29细胞为突变型,Lovo细胞为野生型。p53转录表达水平的差异可能与其细胞分化程度及转移有关。

AIM To investigate the correlation between biological characteristics and mRNA transcriptions, protein-expressions of p53 gene in colorectal cancer HT-29 and Lovo cell lines.METHODS The comparative studies on p53 mRNA transcriptional levels were conducted by analysis of Northern blot with vacuum transfer, in situ hybridization (ISH) with biotin-probes. The expressions of p53 protein were synchronously studied by immunohistochemistry. RESULTS Northern blot was identical to ISH. The positive p53 mRNA was detected in both the cells, but was more intensive in HT-29. However, p53 protein was only detected in HT-29, and Lovo cell was negative. CONCLUSION There are two different p53 genes in HT-29, Lovo cells. The former is mutant type and the latter is wild type. There may be a correlation between the variations of p53 gene transcription-expressions, cell differentiation and transfer character.