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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。
Objective To investigate the relationship between transcription and protein expression of p53 mRNA and its molecular biological characteristics in HT-29 and Lovo colorectal cancer cells. METHODS Northern blot analysis was performed by RNA vacuum transfer method. In situ hybridization was performed with a photobiotin-labeled probe. The transcription levels of p53 mRNA in these two cells were compared and studied. The immunocytochemical staining method was used to compare the expression of P53 protein in these two kinds of cells. Results Northern blot was consistent with in situ hybridization. Both cells were positive for p53 mRNA transcription, and the former was significantly higher in intensity than the latter. P53 protein was only detected in HT-29 cells and Lovo cells were negative. Conclusion There are different types of p53 gene in both cells, HT-29 cells are mutant and Lovo cells are wild type. The difference in the expression of p53 transcription may be related to the degree of cell differentiation and metastasis.