Impaired BAX protein expression in breast cancer: mutational analysis of the BAX and the p53 gene.
We have previously shown that the pro-apoptotic BAX protein is differentially expressed in breast cancer and in other epithelial tumors. In this line, a reduced BAX protein expression is a negative prognostic factor in various carcinomas including breast cancer. For p53, a trancriptional activator of BAX in apoptosis, mutations in the coding sequence were shown to modulate BAX protein expression in cell line models on the transcriptional level. We therefore investigated the BAX gene in 68 breast cancer specimens for the presence of mutations in the coding sequence by single-strand conformation polymorphism (SSCP)-PCR and direct sequencing. The expression of BAX protein was assessed by immunohistochemistry. In addition, we screened for mutations in the exons 5-8 of the p53 gene by SSCP-PCR to assess whether mutations in the DNA-binding domain of this upstream regulator of BAX gene transcription are responsible for differences in BAX protein expression. As previously observed, BAX was differentially expressed in the breast cancer samples, but no mutations in the coding sequence of the BAX gene were found besides a polymorphism in exon 6 at the position 552 (G->A) and additional intronic polymorphisms. In contrast, we identified 16 of 68 (23.5%) tumors to bear mutations in the p53 gene. In the subset of BAX-expressing tumors, the mutational inactivation of p53 did result in a reduced BAX protein expression (Fisher exact test, p = 0. 047). Nevertheless, we identified a subset of BAX-negative tumors lacking BAX or p53 mutations. Thus, additional, not yet identified regulators, apart from p53, appear to be involved in the regulation of BAX protein expression.
Sturm
I
I
Department of Hematology, Oncology and Tumor Immunology, Humboldt University, Berlin-Buch, Germany.
Papadopoulos
S
S
Hillebrand
T
T
Benter
T
T
Lück
H J
HJ
Wolff
G
G
Dörken
B
B
Daniel
P T
PT
D001483:Base Sequence; D001665:Binding Sites; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis* / Q000235:genetics*; D019253:Proto-Oncogene Proteins c-bcl-2; D011960:Receptors, Estrogen / Q000378:metabolism; D011980:Receptors, Progesterone / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; D004273:DNA, Neoplasm; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D051028:bcl-2-Associated X Protein