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

Mühlbauer, Karl-Rudolf; Gröne, Hermann–Josef; Ernst, Thomas; Gröne, Elisabeth F.; Tschada, R.; Hergenhahn, Manfred; Hollstein, Monica

doi:10.1038/sj.bjc.6601164

Cited by 41 publications

(29 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Narla et al 9 originally found KLF6 mutations in 18 of 33 (55%) prostate carcinomas. More recently, Chen et al 12 detected somatic sequence changes in only 14 of 96 prostate cancer samples (15%) whereas Mühlbauer et al 15 did not identify any KLF6 mutations in 32 prostate cancers.…”

Section: Dear Sirmentioning

confidence: 99%

Absence of mutations in the putative tumor suppressor gene KLF6 in glioblastomas and meningiomas

Köhler

Wolter

Blaschke

et al. 2004

Intl Journal of Cancer

View full text Add to dashboard Cite

Section: Dear Sirmentioning

confidence: 99%

Absence of mutations in the putative tumor suppressor gene KLF6 in glioblastomas and meningiomas

Köhler

Wolter

Blaschke

et al. 2004

Intl Journal of Cancer

View full text Add to dashboard Cite

“…3 A comparable frequency of deletions on 10p has been found in prostate cancer. 4 KLF6, a gene that is part of the Kruppel family of transcription factors, 5 is located on 10p and according to several reports is mutated in about 50% of these patients 6 (a recent report, however, does not confirm this finding 7 ).…”

Section: Dear Sirmentioning

confidence: 99%

KLF6 is not the major target of chromosome 10p losses in glioblastomas

Montanini

Bissola

Finocchiaro

2004

Intl Journal of Cancer

View full text Add to dashboard Cite

Dear Sir,Complete or partial deletions of chromosome 10 are critical events during the malignant progression of gliomas. 1 They are present in the large majority of patients with glioblastoma multiforme (GBM), the most malignant glioma. Recent evidence suggests that complete losses of one copy of chromosome 10 are more frequent in primary GBM and partial losses in secondary GBM. 2 More often deletions affect the long arm of chromosome 10 but in a large fraction of patients they are also present on the short arm. 3 A comparable frequency of deletions on 10p has been found in prostate cancer. 4 KLF6, a gene that is part of the Kruppel family of transcription factors, 5 is located on 10p and according to several reports is mutated in about 50% of these patients 6 (a recent report, however, does not confirm this finding 7 ).A recent report by Jeng and Hsu describes the presence of KLF6 mutations in astrocytic gliomas and concludes that "mutations of the KLF6 gene play a role in the pathogenesis of astrocytic gliomas." 8 We have investigated the loss of heterozygosity in the chromosomal region flanking KLF6 and looked for mutation of KLF6 in 34 patients with GBM and in 2 GBM cell lines. LOH studies were done using microsatellite markers KLF6-M1 and KLF6-M2 and methods described previously. 9 LOH with primer M1 (telomeric) was present in 16/34 cases (47%). LOH with primer M2 (centromeric) was found in 8/34 (24%). Overall at least one loss was present in 62% of the cases but only in 3 tumors allelic imbalance was present on both the markers flanking KLF6.We have analyzed the coding sequence of KLF6 in these 34 patients. The 4 exons were amplified using PCR primers and temperatures outlined in Table I.The sequence was found identical to normal in all patients, with one exception. In Patient 19, showing LOH on M1, exons 1, 3 and 4 could be amplified by PCR whereas exon 2 failed to be amplified. Using PCR primers on exons 1 and 4 we could amplify more than 5 kb of sequence of KLF6 in 2 independent DNAs but not in tumor DNA of Patient 19, suggesting the presence of a translocation interrupting the coding sequence of KLF6 in exon 2. In Patient 8, the sequence of lymphocyte DNA showed heterozygosity on codon 201 (CGG/CGA) and loss of heterozygosity on tumor DNA (CGA). Both codons however encode the same arginine residue (reverse sequences are shown in the Fig. 1).To test whether reduced expression of KLF6 was present in some patients, we analyzed by real-time PCR the expression of KLF6 in 8 patients, in 2 established cell lines deriving from malignant gliomas (U87 and U373) and in normal brain. RNA was extract with Eurozol (EuroClone, Wetherby, UK) and 1 g of RNA was reverse-transcribed with Expand Reverse Transcriptase (Roche Diagnostics, Milan, Italy). Real-time PCR was carried out using GeneAmp 5700 and TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA) and primers of KLF6 and GAPDH (Assay on Demand cat no. Hs00154550 and Hs99999905, respectively), all from Applied Biosystems. PCR conditions were 2 min ...

show abstract

“…[10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29] In prostate cancer, four studies have analyzed the frequency of KLF6 mutations. 6,[30][31][32] In the first published study, Narla et al 6 showed a very high frequency, 55%, whereas in the following study, Chen et al 30 obtained a lower frequency, around 15%. The results of two recent reports provided frequencies of 0 and 1%.…”

mentioning

confidence: 97%

“…The results of two recent reports provided frequencies of 0 and 1%. 31,32 It should be noted that there were considerable differences in both the selected cohorts and the methodological approaches of these studies, which could, at least in part, explain the discrepancies in their results.…”

mentioning

confidence: 98%

See 1 more Smart Citation

KLF6 and TP53 mutations are a rare event in prostate cancer: distinguishing between Taq polymerase artifacts and true mutations

et al. 2008

View full text Add to dashboard Cite

Krü ppel-like factor 6 (KLF6) has been reported to act as a tumor suppressor gene involved in the regulation of the cell cycle by activating p21 in a p53-independent manner. Many studies suggest that KLF6 is inactivated by allelic loss and somatic mutation. However, there is a high variability in the reported frequency of mutations (from 1 to 55%). TP53 also regulates the cell cycle through the activation of p21. In prostate cancer, the reported frequency of TP53 mutations ranges from 3 to 42%. In all these reports, there is a considerable degree of methodological heterogeneity. Our aim was to determine the frequency of KLF6 and TP53 mutations in a welldefined group of prostate tumors with different stages and Gleason grades. The four exons of KLF6 and exons 4-9 of TP53 were studied in 103 cases, including 90 formalin-fixed, paraffin-embedded (FFPE) and 13 frozen samples. All tumors were analyzed through PCR and direct sequencing. All changes found were confirmed by a second independent PCR and sequencing reaction. For KLF6, mutation (E227G) was only detected in one tumor (1%) and for TP53, three different mutations (L130H, H214R, and Y234C) were detected in five tumors (5%). This low mutation index is in keeping with recent papers on the subject. Our study strongly supports the notion that KLF6 and TP53 mutations are not frequent events in prostate cancer. When using FFPE tissues, it is mandatory to perform at least two independent rounds of PCR and sequencing to confirm mutations and exclude Taq polymerase-induced artifacts. Two of these genes are Krü ppel-like factor 6 (KLF6) and TP53.KLF6 is a transcription factor that interacts with DNA through three zinc-fingers in its COOHterminal domain. KLF6 belongs to the KLF family, a family that is broadly involved in cell differentiation, development, growth-related signal transduction, cell proliferation, apoptosis, and angiogenesis. 5 It has been suggested that KLF6 could be involved in the regulation of the cell cycle by activating p21 in a p53-independent manner. 6 This has been proven in several in vitro 5 and in vivo assays. 7 KLF6 is believed to regulate cancer development and progression through the downregulation of the c-Jun oncoprotein 8 and the activation of E-cadherin. 9 The reported frequency of KLF6 mutations in different types of human cancer varies in the different published studies. [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29] In prostate cancer, four studies have analyzed the frequency of KLF6 mutations. 6,[30][31][32] In the first published study, Narla et al 6 showed a very high frequency, 55%, whereas in the following study, Chen et al 30 obtained a lower frequency, around 15%. The results of two recent reports provided frequencies of 0 and 1%. 31,32 It should be noted that there were considerable

show abstract

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

Cited by 41 publications

References 11 publications

Absence of mutations in the putative tumor suppressor gene KLF6 in glioblastomas and meningiomas

Absence of mutations in the putative tumor suppressor gene KLF6 in glioblastomas and meningiomas

KLF6 is not the major target of chromosome 10p losses in glioblastomas

KLF6 and TP53 mutations are a rare event in prostate cancer: distinguishing between Taq polymerase artifacts and true mutations

Contact Info

Product

Resources

About