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BRCA1/2 mutation screening and LOH analysis of lung adenocarcinoma tissue in a multiple-cancer patient with a strong family history of breast cancer.

Boettger MB, Sergi C, Meyer P - J Carcinog (2003)

Bottom Line: Using the laser-capture microdissection (LCM) technique, we obtained pure populations of neoplastic cells from which DNA could be extracted.CONCLUSION: To our knowledge, this is the first report of investigation for LOH for BRCA2 in primary lung adenocarcinoma tissue of a patient with multiple primary tumours related to a familial germline BRCA2 mutation.Interestingly, it was the mutant, not the wild-type, allele which was lost in the lung adenocarcinoma tissue.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Human Genetics, University of Heidelberg, 69120 Heidelberg, Germany. Peter.Meyer@onkogenetik.de

ABSTRACT
BACKGROUND: Germline mutations in BRCA1/2 greatly elevate risks of breast and ovarian cancers, but the role of these genes in tumourigenesis of other cancer types is still being investigated. OBJECTIVE: We report on an investigation of BRCA1/2 mutations and their loss of heterozygosity (LOH) in a patient with a strong family history of breast cancer who was diagnosed with consecutive primary cervical, ovarian and lung carcinomas. METHODS AND RESULTS: BRCA1/2 mutation screening of the proband revealed a common familial breast- and ovarian cancer-associated germline BRCA2 mutation (3034del4bp). We then performed LOH analysis for BRCA2 in lung adenocarcinoma tissue of the patient. Using the laser-capture microdissection (LCM) technique, we obtained pure populations of neoplastic cells from which DNA could be extracted. Mutation analysis by denaturing high-performance liquid chromatography (DHPLC) and direct sequencing revealed loss of the mutant allele in the adenocarcinoma tumour tissue. CONCLUSION: To our knowledge, this is the first report of investigation for LOH for BRCA2 in primary lung adenocarcinoma tissue of a patient with multiple primary tumours related to a familial germline BRCA2 mutation. Interestingly, it was the mutant, not the wild-type, allele which was lost in the lung adenocarcinoma tissue.

No MeSH data available.


Related in: MedlinePlus

DHPLC elution profiles of 463 base pair PCR fragments of BRCA2 exon 11. A: Heteroduplex profile with two peaks from control DNA that was found to be heterozygous for the common exon 11 BRCA2 mutation 3034del4bp. B: Homoduplex profile (DHPLC) in amplified wild-type DNA. C: DHPLC analysis of amplified genomic DNA of our patient showing a heteroduplex profile similar to the profile of the mutant control DNA sample. D: DHPLC elution profile of amplified lung tumour DNA showing only one peak indicating LOH, either of the wild-type or the mutant allele.
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Figure 2: DHPLC elution profiles of 463 base pair PCR fragments of BRCA2 exon 11. A: Heteroduplex profile with two peaks from control DNA that was found to be heterozygous for the common exon 11 BRCA2 mutation 3034del4bp. B: Homoduplex profile (DHPLC) in amplified wild-type DNA. C: DHPLC analysis of amplified genomic DNA of our patient showing a heteroduplex profile similar to the profile of the mutant control DNA sample. D: DHPLC elution profile of amplified lung tumour DNA showing only one peak indicating LOH, either of the wild-type or the mutant allele.

Mentions: We obtained prior written informed consent from our patient using protocols approved by the Ethics Committee of the Medical Faculty at the University of Heidelberg, Germany. Genomic DNA was isolated from blood using standard procedures. To amplify exon and exon-intron boundaries, we used primer pairs and polymerase chain reaction (PCR) protocols as previously published [5]. Mutational screening for BRCA1 and BRCA2 mutations was carried out by prescreening with denaturing high-performance liquid chromatography (DHPLC) analysis and direct sequencing as described elsewhere [5]. DHPLC analysis of amplified genomic DNA of our patient showed a distinctive elution profile in exon 11 of BRCA2 (Fig. 2A,2B,2C), and dye terminator sequencing identified the exact sequence alteration. We detected the common breast cancer-associated frameshift mutation 3034del4bp (Fig. 3A), which causes premature protein termination at codon 958.


BRCA1/2 mutation screening and LOH analysis of lung adenocarcinoma tissue in a multiple-cancer patient with a strong family history of breast cancer.

Boettger MB, Sergi C, Meyer P - J Carcinog (2003)

DHPLC elution profiles of 463 base pair PCR fragments of BRCA2 exon 11. A: Heteroduplex profile with two peaks from control DNA that was found to be heterozygous for the common exon 11 BRCA2 mutation 3034del4bp. B: Homoduplex profile (DHPLC) in amplified wild-type DNA. C: DHPLC analysis of amplified genomic DNA of our patient showing a heteroduplex profile similar to the profile of the mutant control DNA sample. D: DHPLC elution profile of amplified lung tumour DNA showing only one peak indicating LOH, either of the wild-type or the mutant allele.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC239936&req=5

Figure 2: DHPLC elution profiles of 463 base pair PCR fragments of BRCA2 exon 11. A: Heteroduplex profile with two peaks from control DNA that was found to be heterozygous for the common exon 11 BRCA2 mutation 3034del4bp. B: Homoduplex profile (DHPLC) in amplified wild-type DNA. C: DHPLC analysis of amplified genomic DNA of our patient showing a heteroduplex profile similar to the profile of the mutant control DNA sample. D: DHPLC elution profile of amplified lung tumour DNA showing only one peak indicating LOH, either of the wild-type or the mutant allele.
Mentions: We obtained prior written informed consent from our patient using protocols approved by the Ethics Committee of the Medical Faculty at the University of Heidelberg, Germany. Genomic DNA was isolated from blood using standard procedures. To amplify exon and exon-intron boundaries, we used primer pairs and polymerase chain reaction (PCR) protocols as previously published [5]. Mutational screening for BRCA1 and BRCA2 mutations was carried out by prescreening with denaturing high-performance liquid chromatography (DHPLC) analysis and direct sequencing as described elsewhere [5]. DHPLC analysis of amplified genomic DNA of our patient showed a distinctive elution profile in exon 11 of BRCA2 (Fig. 2A,2B,2C), and dye terminator sequencing identified the exact sequence alteration. We detected the common breast cancer-associated frameshift mutation 3034del4bp (Fig. 3A), which causes premature protein termination at codon 958.

Bottom Line: Using the laser-capture microdissection (LCM) technique, we obtained pure populations of neoplastic cells from which DNA could be extracted.CONCLUSION: To our knowledge, this is the first report of investigation for LOH for BRCA2 in primary lung adenocarcinoma tissue of a patient with multiple primary tumours related to a familial germline BRCA2 mutation.Interestingly, it was the mutant, not the wild-type, allele which was lost in the lung adenocarcinoma tissue.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Human Genetics, University of Heidelberg, 69120 Heidelberg, Germany. Peter.Meyer@onkogenetik.de

ABSTRACT
BACKGROUND: Germline mutations in BRCA1/2 greatly elevate risks of breast and ovarian cancers, but the role of these genes in tumourigenesis of other cancer types is still being investigated. OBJECTIVE: We report on an investigation of BRCA1/2 mutations and their loss of heterozygosity (LOH) in a patient with a strong family history of breast cancer who was diagnosed with consecutive primary cervical, ovarian and lung carcinomas. METHODS AND RESULTS: BRCA1/2 mutation screening of the proband revealed a common familial breast- and ovarian cancer-associated germline BRCA2 mutation (3034del4bp). We then performed LOH analysis for BRCA2 in lung adenocarcinoma tissue of the patient. Using the laser-capture microdissection (LCM) technique, we obtained pure populations of neoplastic cells from which DNA could be extracted. Mutation analysis by denaturing high-performance liquid chromatography (DHPLC) and direct sequencing revealed loss of the mutant allele in the adenocarcinoma tumour tissue. CONCLUSION: To our knowledge, this is the first report of investigation for LOH for BRCA2 in primary lung adenocarcinoma tissue of a patient with multiple primary tumours related to a familial germline BRCA2 mutation. Interestingly, it was the mutant, not the wild-type, allele which was lost in the lung adenocarcinoma tissue.

No MeSH data available.


Related in: MedlinePlus