The p53 codon 249 mutant--derived from human functional adrenal tumors--can modify the cell shape of normal adrenocortical transfected cells.
Our previous study has indicated that p53 gene mutation occurred in 73% of human functional adrenal tumors, and the mutation hot spots were focused on codons 100, 102 (exon 4), and 249 (exon 7). Furthermore, a transcriptional activity study revealed that the mutant p53 protein derived from human functional adrenal tumors lost 90% transcriptional activity and the ability to bind with the p53 sequence. In order to investigate the influence of the mutant protein extracted from adrenocortical tumors on normal adrenal cells, we first cloned p53 cDNA from the human primary aldosteronism and constituted it with isopropyl thiogalactoside (IPTG) inducible expression vector as recombinant plasmid. The recombinant plasmid was then transfected to normal bovine adrenocortical cells through electroporation. The results showed that the p53 protein mutations at codons 100 and 102 could neither affect the cell morphology nor enable cell growth on the soft agar. In addition, no significant difference was found in cortisol level between the p53 transfected and the control cells. On the other hand, cell morphological changes and cell proliferation rate increase were observed when we used IPTG to induce the expression of the p53 protein, which mutated at codon 249, in adrenocortical cells. The cell morphology changes included less flattened and decreased elongation when compared to non-transfected cells. However, the cortisol level in transfected cells was not affected by the p53 mutants. Taken together, we concluded that the mutant p53 protein indeed participates in adrenal carcinogenesis; however, it has no influence on hormone production and secretion.
Hsu
C H
CH
Department of Clinical Pathology, Kaohsiung Medical University, No. 100, Shih-Chuan 1st Road, 80317, Kaohsiung, Taiwan, ROC.
Lee
S C
SC
Yang
Y C
YC
Lian
S T
ST
Shin
S J
SJ
Lin
S R
SR
D000302:Adrenal Cortex / Q000473:pathology; D000310:Adrenal Gland Neoplasms / Q000235:genetics* / Q000473:pathology; D048429:Cell Size / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003062:Codon; D016158:Genes, p53; D006801:Humans; D009154:Mutation; D014162:Transfection; D014407:Tumor Cells, Cultured
D003062:Codon