Truncating mutations of PPM1D are found in blood DNA samples of lung cancer patients.
BACKGROUND
PPM1D (WIP1) negatively regulates by dephosphorylation many proteins including p53 tumour suppressor. The truncating mutations (nonsense and frameshift) in exon 6 of PPM1D were found recently in blood cells of patients with breast, ovarian or colorectal cancer. These mutants code for gain-of-function PPM1D with retained phosphatase activity. Their significance in carcinogenesis is unknown.


METHODS
The exon 6 of PPM1D was sequenced in blood DNA of 543 non-small-cell lung cancer patients (NSCLC). The functional significance of selected PPM1D alterations (Arg458X, Lys469Glu) was compared with the wild-type gene and examined by recombinant DNA techniques, immunoblotting and luciferase reporter assays.


RESULTS
The frameshift mutations were found in five NSCLC patients (5/543; 0.92%), all of them had squamous cell carcinomas (5/328; 1.5%). All patients with the mutations were exposed, before the blood collection, to the DNA damaging agents as a part of chemotherapeutic regimen. Functional tests demonstrated that truncating mutation Arg458X causes enhancement of dephosphorylation activity of PPM1D toward serine 15 of p53, whereas Lys469Glu version is equivalent to the wild-type. Neither version of PPM1D (wild-type, Arg458X, Lys469Glu) significantly modulated the ability of p53 to transactivate promoters of the examined p53-target genes (BAX and MDM2).


CONCLUSIONS
The truncating mutations of PPM1D are present in blood DNA of NSCLC patients at frequency similar to percentage determined for ovarian cancer patients. Our findings raise a question if the detected lesions are a result of chemotherapy.
Center for Translational Research and Molecular Biology of Cancer, Maria Skłodowska -Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland.
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Zajkowicz
Center for Translational Research and Molecular Biology of Cancer, Maria Skłodowska -Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland.
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Butkiewicz
1] II Clinic of Radiotherapy and Chemotherapy, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland [2] Department of Clinical Oncology, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland.
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Drosik
II Clinic of Radiotherapy and Chemotherapy, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland.
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Giglok
II Clinic of Radiotherapy and Chemotherapy, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland.
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Suwiński
Center for Translational Research and Molecular Biology of Cancer, Maria Skłodowska -Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, 44-101 Gliwice, Poland.
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Rusin
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002289:Carcinoma, Non-Small-Cell Lung / Q000097:blood / Q000235:genetics; D018389:Codon, Nonsense; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000097:blood / Q000235:genetics*; D005091:Exons; D005260:Female; D016368:Frameshift Mutation; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D010749:Phosphoprotein Phosphatases / Q000097:blood* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D000071636:Protein Phosphatase 2C; D012694:Serine / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D055815:Young Adult
D018389:Codon, Nonsense; D004273:DNA, Neoplasm; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D012694:Serine; C000607458:PPM1D protein, human; D010749:Phosphoprotein Phosphatases; D000071636:Protein Phosphatase 2C