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Analysis of human prostate cancers and cell lines for mutations in the TP53 and KLF6 tumour suppressor genes.

Mühlbauer KR, Gröne HJ, Ernst T, Gröne E, Tschada R, Hergenhahn M, Hollstein M - Br. J. Cancer (2003)

Bottom Line: The aims of the present study were to confirm these initial findings by determining the frequency of exon2 KLF6 mutations in a cohort of European prostate cancer patients, and to investigate whether there was evidence for mutational inactivation of both the KLF6 and TP53 tumour suppressor loci in some tumours.We examined 32 primary prostate tumours and three prostate tumour cell lines for mutations by PCR amplification and direct dideoxy sequencing (KLF6), and by oligonucleotide microarray (p53GeneChip) analysis and dideoxy sequencing (TP53).Whereas TP53 mutations typical of prostate cancer were found at a frequency consistent with the literature, no KLF6 mutations were found in any of the tumour samples nor in the three prostate cancer cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Alterations in Carcinogenesis, German Cancer Research Center (Deutsches Krebsforschungszentrum), D-69120 Heidelberg, Germany.

ABSTRACT
A recent report suggests that the KLF6 gene encoding the Krüppel-like factor 6 protein is a frequently mutated, putative tumour suppressor gene in prostate cancer. The aims of the present study were to confirm these initial findings by determining the frequency of exon2 KLF6 mutations in a cohort of European prostate cancer patients, and to investigate whether there was evidence for mutational inactivation of both the KLF6 and TP53 tumour suppressor loci in some tumours. We examined 32 primary prostate tumours and three prostate tumour cell lines for mutations by PCR amplification and direct dideoxy sequencing (KLF6), and by oligonucleotide microarray (p53GeneChip) analysis and dideoxy sequencing (TP53). Whereas TP53 mutations typical of prostate cancer were found at a frequency consistent with the literature, no KLF6 mutations were found in any of the tumour samples nor in the three prostate cancer cell lines.

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TP53 analysis of tumour 2 by P53GeneChip™ and by Sanger dideoxy sequencing. (A) Bar chart shows signal intensities from array tiling that screened codon 190 from a P53GeneChip hybridised with target from tumour 2. The C to T transition at codon 190 is indicated by the bar at position 2 [C C T to C T/C T, Proline (P) to Leucine (L)]. (B) Electropherogram from ABI Genetic Analyzer showing p53 exon 6 DNA sequence (5′ to 3′) in tumour 2. The ‘N’ shows the position of the C to T transition at the second position of codon 190.
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fig1: TP53 analysis of tumour 2 by P53GeneChip™ and by Sanger dideoxy sequencing. (A) Bar chart shows signal intensities from array tiling that screened codon 190 from a P53GeneChip hybridised with target from tumour 2. The C to T transition at codon 190 is indicated by the bar at position 2 [C C T to C T/C T, Proline (P) to Leucine (L)]. (B) Electropherogram from ABI Genetic Analyzer showing p53 exon 6 DNA sequence (5′ to 3′) in tumour 2. The ‘N’ shows the position of the C to T transition at the second position of codon 190.

Mentions: We did not find mutations in exon 2 of the KLF6 gene in any of the 32 tumour samples that we examined (Table 1) nor in the human prostate tumour cell lines PC-3, DU-145 and LNCaP. The P53GeneChip™ protocol identified TP53 gene mutations in three of the 25 tumours (12%). All were transitions at G : C base pairs (the most common type of mutation in this cancer type) at codons previously reported mutated in prostate and other cancers (Olivier et al, 2002). The presence of these mutations was confirmed by amplifying genomic DNA with p53-specific primers, and then performing cycle sequencing and electrophoretic analysis following the same standard methodology used for KLF6 analysis (Table 1, Figure 1Figure 1


Analysis of human prostate cancers and cell lines for mutations in the TP53 and KLF6 tumour suppressor genes.

Mühlbauer KR, Gröne HJ, Ernst T, Gröne E, Tschada R, Hergenhahn M, Hollstein M - Br. J. Cancer (2003)

TP53 analysis of tumour 2 by P53GeneChip™ and by Sanger dideoxy sequencing. (A) Bar chart shows signal intensities from array tiling that screened codon 190 from a P53GeneChip hybridised with target from tumour 2. The C to T transition at codon 190 is indicated by the bar at position 2 [C C T to C T/C T, Proline (P) to Leucine (L)]. (B) Electropherogram from ABI Genetic Analyzer showing p53 exon 6 DNA sequence (5′ to 3′) in tumour 2. The ‘N’ shows the position of the C to T transition at the second position of codon 190.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376923&req=5

fig1: TP53 analysis of tumour 2 by P53GeneChip™ and by Sanger dideoxy sequencing. (A) Bar chart shows signal intensities from array tiling that screened codon 190 from a P53GeneChip hybridised with target from tumour 2. The C to T transition at codon 190 is indicated by the bar at position 2 [C C T to C T/C T, Proline (P) to Leucine (L)]. (B) Electropherogram from ABI Genetic Analyzer showing p53 exon 6 DNA sequence (5′ to 3′) in tumour 2. The ‘N’ shows the position of the C to T transition at the second position of codon 190.
Mentions: We did not find mutations in exon 2 of the KLF6 gene in any of the 32 tumour samples that we examined (Table 1) nor in the human prostate tumour cell lines PC-3, DU-145 and LNCaP. The P53GeneChip™ protocol identified TP53 gene mutations in three of the 25 tumours (12%). All were transitions at G : C base pairs (the most common type of mutation in this cancer type) at codons previously reported mutated in prostate and other cancers (Olivier et al, 2002). The presence of these mutations was confirmed by amplifying genomic DNA with p53-specific primers, and then performing cycle sequencing and electrophoretic analysis following the same standard methodology used for KLF6 analysis (Table 1, Figure 1Figure 1

Bottom Line: The aims of the present study were to confirm these initial findings by determining the frequency of exon2 KLF6 mutations in a cohort of European prostate cancer patients, and to investigate whether there was evidence for mutational inactivation of both the KLF6 and TP53 tumour suppressor loci in some tumours.We examined 32 primary prostate tumours and three prostate tumour cell lines for mutations by PCR amplification and direct dideoxy sequencing (KLF6), and by oligonucleotide microarray (p53GeneChip) analysis and dideoxy sequencing (TP53).Whereas TP53 mutations typical of prostate cancer were found at a frequency consistent with the literature, no KLF6 mutations were found in any of the tumour samples nor in the three prostate cancer cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Alterations in Carcinogenesis, German Cancer Research Center (Deutsches Krebsforschungszentrum), D-69120 Heidelberg, Germany.

ABSTRACT
A recent report suggests that the KLF6 gene encoding the Krüppel-like factor 6 protein is a frequently mutated, putative tumour suppressor gene in prostate cancer. The aims of the present study were to confirm these initial findings by determining the frequency of exon2 KLF6 mutations in a cohort of European prostate cancer patients, and to investigate whether there was evidence for mutational inactivation of both the KLF6 and TP53 tumour suppressor loci in some tumours. We examined 32 primary prostate tumours and three prostate tumour cell lines for mutations by PCR amplification and direct dideoxy sequencing (KLF6), and by oligonucleotide microarray (p53GeneChip) analysis and dideoxy sequencing (TP53). Whereas TP53 mutations typical of prostate cancer were found at a frequency consistent with the literature, no KLF6 mutations were found in any of the tumour samples nor in the three prostate cancer cell lines.

Show MeSH
Related in: MedlinePlus