Mobile website
Detection of P53 gene mutations in 14 tumor cell lines
- Article
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
Abstract:
To investigate P53 gene mutations in 14 tumor cell lines. Methods: Primers were designed according to the intron sequences locating between exons 5 8 of P53 gene for amplification of P53 gene exon 5 8 of the 14 tumor cell lines from 9 kinds of human carcinoma, namely, the lung cancer, gloma, hepatocarcinoma, gastric carcinoma, prostate carcinoma, mammary carcinoma, colonic carcinoma, choroidal melanoma, and retinoblastoma. The PCR products were subjected to electrophoresis; DNA sequencing was performed for the amplified product and the results were compared with wild type P53 gene. Meanwhile, the P53 protein of 14 tumor cell lines was extracted for Western blotting analysis. Results: The products of exons 5 8 amplified by PCR were identical to that expected. The results of DNA sequencing showed that 8 of 14 tumor cell lines had P53 gene mutation in exons 5 8. Novel mutations were found in 4 tumor cell lines, including human lung cancer cell H1299, hepatocacinoma cell Hep3B and 7721, and choroidal melanoma cell OCM 1. The mutations mainly existed in the coding areas of the exons; most of them were missense mutations due to single base replacement; some others were silent mutations; and no mutations were found in the 2 normal cell lines. The results of Western blotting showed that only 6 of 8 mutant lines had P53 protein expression, which was not found in the non mutant lines and the normal cells. Conclusion: We have identified 8 lines with mutant P53 genes among the 14 tumor cell lines, novel mutations are found in 4 tumor cell lines and none in the two normal cell lines.
Keywords:
tumor cell ; P53 gene ; mutation
Project Supported:
Supported by the National Natural Science Foundation of China（No.39770242）
