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[Cancer Research 60, 13-17, January 1, 2000]
© 2000 American Association for Cancer Research

Advances in Brief

Molecular Cloning and Characterization of Human MAWD, a Novel Protein Containing WD-40 Repeats Frequently Overexpressed in Breast Cancer1

Satoru Matsuda2, Ryu Katsumata, Takahito Okuda, Tatsuyoshi Yamamoto, Kou Miyazaki, Takeshi Senga, Kazuya Machida, Aye Aye Thant, Shigekazu Nakatsugawa and Michinari Hamaguchi

Department of Molecular Pathogenesis [S. M., R. K., T. O., K. M., T. S., A. A. T., M. H.] and First Department of Surgery [T. Y., K. M.], Nagoya University School of Medicine, Nagoya 466-8550, Japan, and Division of Radiology, Nagoya University, Daiko Medical Center, Nagoya 461-0047 Japan [S. N.]


   ABSTRACT
Top
 ABSTRACT
Introduction
 Materials and Methods
 Results and Discussion
 REFERENCES
 
A full-length cDNA clone encoding a novel protein containing WD-40 repeats, which were frequently involved in protein-protein interactions, was isolated and sequenced. This clone had a predicted open reading frame (ORF) encoding 350 amino acids possessing six repeats of WD-40 motif. It was most closely homologous to TRIP-1, a phosphorylation substrate of the transforming growth factor-ß type II receptor. In the process of characterizing the function of the new gene product, we found that overexpression of the gene seemed to activate mitogen-activated protein kinase and to promote anchorage-independent growth of the cells. Moreover, the gene product was frequently overexpressed in human tumor breast tissues compared with their normal breast tissues, suggesting that the gene might be involved in the tumor progression. Radiation hybrid mapping placed the gene into human chromosome 12q11–12 near the marker D12S1593.


   Introduction
Top
 ABSTRACT
 Introduction
Materials and Methods
 Results and Discussion
 REFERENCES
 
Accumulating evidence suggests that a cancer arises through accumulation of a number of genetic lesions. In addition, loss of sensitivity to negative growth regulators is thought to be an important step in the development of neoplastic lesions. TGF3 -ß is one of the most elucidated negative growth regulators of cells. However, the exact mechanism(s) for signaling from the receptors for TGF-ß remain to be further elucidated. To date, several types of signaling molecules have been identified as downstream mediators of the TGF-ß receptors. Among them, TRIP-1 has unique function and structure containing WD-40 motif (1) .

The WD repeats proteins are characterized by a cluster of repeated sequences, each repeating around 40 amino acids in length and usually ending with tryptophan-aspartate (WD), hence the name. The WD-40 repeats are found in proteins involved in a wide variety of cellular processes ranging from signal transduction to RNA processing (2) . Proteins containing WD repeats are often physically associated with other proteins and are believed in many cases to act as scaffolds upon which the multimeric complexes are built (3) . The crystal structure of the heterotrimeric G protein revealed that the WD repeats of the subunit fold into a highly symmetrical, propeller-like structure with the conserved region of each repeat forming a part of the propeller blade reviewed by Neer and Schmidt (4) .

These exciting findings prompted us to try to discover a new WD-40 repeats protein that might be involved in cell growth or cancer progression. In the present study, we report the cloning and sequencing of a full-length cDNA that codes for a novel WD-40 repeats protein similar to TRIP-1. As described later, we name the gene product hMAWD, standing for a putative human MAPK activator with WD repeats. The possible roles of the gene are also discussed.


   Materials and Methods
Top
 ABSTRACT
 Introduction
 Materials and Methods
Results and Discussion
 REFERENCES
 
Identification of the Expressed Sequence Tag Sequences.
To find a new member of the human WD-40 repeats protein, the WD-40 motifs of TRIP-1 were used to search homologues in gene data bases stored at the National Center for Biotechnology by using the tBLASTn sequence alignment program. w313 cDNA, which was identified by the similarity search, was amplified by RT-PCR procedure from the mRNA of human normal tissues.

Screening of cDNA and Sequencing.
A human HepG2 cDNA library constructed using lambda ZAP vector (Stratagene) was purchased and screened by using standard conditions. Briefly, 1 x 106 clones in total were screened with the ECL-labeled cDNA fragment. Positive clones were selected, and their insert cDNAs were excised in vivo in pBluescript II, following the supplier’s recommendation. The longest cDNA thus constructed was sequenced to completion of both strands using the Thermo Sequenase Dye Termination Cycle Sequencing pre-mix kit (Amersham) on an A.L.F. DNA sequencer II (Pharmacia LKB). The search for the sequence analysis and the comparison to related sequences were carried out using tBLASTn program.

Chromosomal Assignment.
Chromosomal assignment of the hMAWD gene was accomplished by PCR analysis of genomic DNAs of the Genebridge 4 RH panel (Research Genetics, Huntsville, AL) as well as those of Stanford G3 RH panel (Research Genetics). The Ums (sense, 5'-TGAATTAGCTCCAGTG) and Dma (antisense, 5'-ATAACAGGCCACTGTA) PCR primers were designed specifically to amplify the hMAWD sequence in the rodent background genes. PCR consisted of 35 cycles (94°C for 40 s, 56°C for 40 s, and 72°C for 40 s) after the initial denaturation step (94°C for 8 min). The data for hMAWD of Genebridge 4 RH panel were as follows: 111111111000111101011010000100001011111111000110111110010010-0111111110011101111101111.

Cell Culture.
COS-7 cells, ECV304 cells, and the derived cells were cultured in DMEM with 10% fetal bovine serum and antibiotics.

Colony Formation in Soft Agar.
Cells (1 x 104) were suspended in 2 ml of 0.4% agar in DMEM containing 10% FBS and overlayed on a layer of 0.8% agar in medium. After 10 days, the number of colonies <1.0 mm in diameter was counted.

Human Tissue Samples.
Fresh surgical specimens of 46 patients diagnosed histologically as having breast cancer (46 cases of invasive ductal adenocarcinoma) and their adjacent normal tissues were obtained from patients undergoing surgery. The materials, obtained immediately after the surgical procedure, were frozen in liquid nitrogen and stored at -80°C.

Development of Specific Antibody.
For the purpose of producing specific antibodies, the partial hMAWD (amino acids 173–350) and the full-length hMAWD were expressed and purified as GST fusion proteins using bacterial expression vector pGEX5X-1 (Pharmacia Biotech, Inc.). Rabbits were immunized, and specific antibody was prepared using the fusion protein.

Immunofluorescence Analysis and Immunohistochemistry.
These methods were basically performed as described previously (5 , 6) . Then, the cells were inspected with a confocal scanning microscopy and standard microscopy.

Western Blot Analysis.
Total cell lysates were prepared by suspending 100-mg samples in RIPA buffer [40 mM Tris-HCl (pH 7.5), 1% Triton X-100, and 150 mM NaCl]. The total cell lysate was resuspending in a standard Laemmli sample preparation buffer and loaded onto a 10% polyacrylamide gel. Proteins were immunoblotted to Immobilon filters, and the filters were incubated sequentially with anti-hMAWD polyclonal antibody, alkalyphosphatase-conjugated antirabbit IgG, and developed with the Promega NBT-BCIP detection system suggested in the manufacturer’s instructional protocol.


   Results and Discussion
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results and Discussion
REFERENCES
 
To determine whether there might be additional members of the human WD-40 repeat protein family lurking in one of the sequence databases, the tBLASTn program (7) was used to query each database with the amino acid sequences of the human TRIP-1 protein. The expressed sequence tag database revealed several clones, apparently from the same gene, that shared significant amino acid sequence similarities to the WD-40 repeat motif of TRIP-1. The 300-bp cDNA fragment was amplified by RT-PCR from a human placenta cDNA library and identified as a clone (w313) that has significant homology to TRIP-1. The w313 fragment was then used as a probe for the screening of a full-length cDNA against a human HepG2 cDNA library. Using a photo-labeled w313 probe, a total of four putative positive clones were isolated after screening about 1 x 106 plaques of the cDNA library (8 , 9) . Sequence analysis indicated that all of these clones contain cDNAs from the same encoding a full-length corresponding to w313. The entire sequence of the clones encodes a 1.8-kb nucleotide sequence with a single ORF of 1050 nucleotides. A typical polyadenylation signal (AATAAA) is located downstream from the first in-frame stop codon (10) . The ORF sequence of the clone predicts a protein of 350 amino acids with a calculated molecular weight of Mr 39,000. We applied computer analysis, and the deduced protein sequence had six repeats of WD-40 motif, most closely related to mouse STRAP with 97% amino acid identity over the entire sequence. STRAP is the recently identified novel protein that is associated with TGF-ß receptors from mouse cDNA library by the two-hybrid system (11) . From the alignment and the highly conserved amino acid sequences, the identified cDNA is supposed to be a human homologue of the mouse STRAP gene. To determine the expression patterns of hMAWD mRNA, Northern blot hybridizations and cycle-limited RT-PCR were carried out. It revealed that the hMAWD gene is ubiquitously expressed in all human tissues examined (data not shown). The hMAWD messages are about 2.0 kb, matching the size of the cloned cDNAs. These data are consistent with that of STRAP (11) .

DNA from a panel of rodent-human hybrids, together carrying most human chromosome regions, was tested for presence of the hMAWD gene locus by PCR amplification (12 ; Fig. 1Citation ). Oligonucleotide primers Ums and Dma, corresponding to the 3' untranslated region of the hMAWD gene, were used to amplify specifically hMAWD but not its rodent homologue. To refine the localization of the gene, both of the Genebridge 4 RH panel and the Stanford Gene 3 panel were tested (12) . The results of these data were equal and showed that the hMAWD locus lies near the region to the marker D12S1596 (LOD, 5.5). These findings led to the placement of the hMAWD gene at 12q11–12.



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  		Fig. 1. Representation of the hMAWD cDNA and sequences. A, overlapping hMAWD cDNA clones and a diagram of the complete cDNA structure. The thick line and the striped box represent the untranslated and the coding sequences, respectively. The shaded box indicates the cDNA fragment used as a probe for library screening, and open boxes represent the four overlapping clones. B, cDNA and predicted amino acid sequences in single-letter code of hMAWD. Boldface and roman numbers on the left refer to amino acids and nucleotides, respectively. The hMAWD ORF is from position 301 to 1350. Putative WD-40 motifs are underlined. The presumed polyadenylation site (AATAAA) in the noncoding region of DNA sequence is indicated by a double underline. The nucleotide sequence data reported in this paper have been deposited to DDBJ, EMBL, and GenBank database under the accession number AB024327.

 
To generate cells overexpressing the cDNA for assessing biological effects and for evaluating the function of the hMAWD protein in cell growth, we engineered four different expression vectors containing the full-length untagged, His-tagged, Xpress-tagged, and green fluorescent protein-tagged cDNA in the sense in-frame orientation. These were first tested in transient transfection assays with COS-7 cells and were found to significantly express protein, compared with mock-transfected controls (data not shown). The intracellular localization of both endogenous and overexpressed hMAWD was mainly shown in cytoplasm (Fig. 2, A and BCitation ). Then, to derive stable cell lines overexpressing the protein, we used the well-established human ECV304 cell line (13) . After plasmid gene transfection by Lipofection, drug-resistant colonies were selected and screened. The third panel in Fig. 2CCitation shows a Western blot analysis of the expression in three independent ECV304 clones harboring Xpress-tagged hMAWD sense vector (termed P1, P2, and P3), vector only (C), and parental ECV304 cells (N). All of these cells overexpressing exogenous hMAWD were unexpectedly formed colonies in Petri dishes (data not shown) and exhibited growth in soft agar (Fig. 3, A and B)Citation . Src-transformed cells served as a positive control, and parental ECV304 cells and ECV304 cells harboring vector alone served as negative controls for these assays. These results suggest that targeted up-regulation of hMAWD protein expression disrupts contact inhibition and leads to anchorage-independent growth.



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  		Fig. 2. Cytoplasmic hMAWD staining. A, using a specific anti-His antibody, His-tagged hMAWD was detected in the cytoplasm of the COS-7 cells only transfected with His-tagged full-length hMAWD (top panel). B, using the specific anti-hMAWD antibody, endogenous cytoplasmic hMAWD signals were detected in ECV304 cells (top panel). C, dual-labeling with antibodies directed against hMAWD (top panel) or phospho-MAPK (middle panel) in COS-7 cells transfected with full-length hMAWD. Note that the distribution of phospho-MAPK follows the distribution of hMAWD. The bottom panels in A, B, and C were phase contrasts of the cells. The cells were inspected with a confocal scanning microscopy. D, positive immunoreactivity for hMAWD in the cytoplasm of an invasive ductal breast carcinoma by immunohistological staining. x400.

 


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  		Fig. 3. A, quantitation of cell growth in soft agar is shown. The number of colonies per field is as indicated. The results shown here are the means of quadruplicate determinations and the representative of the three independent experiments; bars, SE. P1, P2, and P3 indicate three independent stable clones overexpressing hMAWD. N and C, parental ECV304 cells and mock vector-transfected ECV304 cells, respectively. Note that the behavior of the clones overexpressing hMAWD is close to that of v-Src-transformed 3Y1 cells (v-Src) under these conditions. B, photograph depicting anchorage-independent colony formation of hMAWD-overexpressed cells (P2) in soft agar. C, stable ECV304 transfectants were analyzed for the expression of the indicated protein. The phospho-MAPK antibody was also used to analyze MAPK activation, showing that hMAWD expression is associated with active MAPK. MAPK monoclonal antibody blot (top panel), phospho-MAPK blot (second panel), Xpress-MAWD blot (third panel), and Tululin blot (bottom panel) as a control are shown.

 
To approach the mechanisms by which overexpression of hMAWD promotes a loss of contact inhibition and anchorage-independent growth, we used a variety of phospho-specific antibodies that have been generated against the activated forms of well-known signal transducer molecules (data not shown). Using antibody detected against activated MAPK among them, we found that MAPK was constitutively activated in all three hMAWD-overexpressed clones (Fig. 3CCitation , second panel). Blotting with MAPK phospho-independent antibody and with Tululin antibody was performed as controls for equal loading (Fig. 3CCitation , top and bottom). Furthermore, it was confirmed by immunostaining that the cells overexpressing hMAWD were just the anti-phospho-MAPK immunoreactive cells (Fig. 2C)Citation . Thus, hMAWD overexpression could lead to constitutive activation of MAPK.

It seems unlikely that activation of MAPK depends on the hMAWD-stimulated autocrine growth factors such as fibroblast growth factor or platelet-derived growth factor because the supernatant of the hMAWD-overexpressing clones could not activate the MAPK of parental cells (data not shown). We have also found that the amount of tyrosine-phosphorylated protein (14) was not increased in hMAWD-overexpressing cells compared with the parental cells (data not shown). Treatment with the specific MAP/extracellular signal-regulated kinase inhibitor (PD98059, 50 mM) reduced the level of activated MAPK to normal levels and blocked the ability to undergo anchorage-independent growth in soft agar; however, ras activation was not found in the hMAWD-overexpressing cells (data not shown). These findings may indicate the Ras-independent MAPK activation by hMAWD. Several Ras-independent pathways have been shown to induce MAPK activation (15, 16, 17) . To gain insight into possible mechanisms of MAPK activation by hMAWD, we are looking for upstream proteins that might interact with hMAWD.

These findings prompted us to investigate whether the hMAWD gene was also overexpressed in human cancers. We found preliminarily that the PCR fragments corresponding to hMAWD mRNA transcripts were well amplified in breast tumors relative to patient-matched normal breast tissues (data not shown). We next evaluated whether the increased expression of hMAWD was paralleled by an increase in immunoreactive protein. Forty-six primary tumor breast tissues that were examined histologically for the presence of tumor cells and patient-matched adjacent normal breast tissues were obtained and examined to quantify the contents in breast carcinomas. None of the patients included in this study had received radiation therapy or chemotherapy before surgery. As shown in Fig. 4Citation , where representative 15 cases were presented, Western blot analyses revealed a immunoreactive band of approximately Mr 39,000 in the tumor tissues that were undetectable in the normal tissue counterpart. In 21 of 46 (45.6%) breast cancer tissues compared with normal tissues, higher expression of hMAWD was detected by immunoblotting analyses. It seemed unlikely that the increases in hMAWD expression in breast cancer tissues merely reflected an increased amount of applied protein, because SHP2, a protein tyrosine phosphatase, expression was not changed when compared with normal tissue (Fig. 4)Citation . Furthermore, by immunohistochemical studies using specific anti-hMAWD antibody, the immunoreactive staining in cytoplasm was observed in the tumor cells (Fig. 2D)Citation . No signal was seen in the normal cells of these specimens, whereas an intensity-variable diffuse cytoplasmic staining in at least 50% of the tumor cells was detected in all breast cancers that had been formerly examined to be hMAWD positive by Western blot analyses, suggesting that the staining was not attributable to fixation artifacts. In this screening approach, hMAWD expression levels were thus independently up-regulated during mammary tumorigenesis.



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  		Fig. 4. Immunoblotting of the lysates from breast cancer tissues and the patient-matched normal tissues. Results of the representative 15 of 46 cases are shown. The tissues were lysed in RIPA buffer, and the lysates (50 mg) were analyzed by immunoblotting with anti-hMAWD antibody as described in "Materials and Methods." All lanes show expression of hMAWD at various levels. P lane, positive control of hMAWD, which is expressed in COS-7 cells transfected with full-length hMAWD. The position of hMAWD protein is indicated by arrowhead on the right side. The SHP2 levels were almost constant throughout, showing that the loading and transfer of the samples were consistent.

 
Breast cancer, which is one of the most common life-threatening diseases in women, occurs in hereditary and sporadic forms. However, because malignant tumor progression is a complex series of sequential steps that incorporate specific properties of tumor cells and host, the mechanism is still not well understood. Our present findings may indicate that the hMAWD gene is a frequent target for alterations in human breast cancers. Similar to the examples of erbB2 or cyclin D, hMAWD may also be useful clinical tumor markers for managing patients with breast cancer (18, 19, 20) . Of course these studies have to be repeated in the future before definitive conclusions can be drawn; however, the novel finding in the present study provides valuable information for understanding the WD-40 repeats proteins and to find the next target of TGF-ß receptor and cancer signals.


   ACKNOWLEDGMENTS
 
We thank Ryoko Miyabe for excellent technical assistance.


   FOOTNOTES
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 This work was supported by a Grant-in-Aid for scientific research on priority areas and for COE Research from the Ministry of Education, Science, Sports and Culture of Japan, a Grant under the Monbusho International Scientific Research Program. The nucleotide sequence data reported in this paper have been deposited to the DNA Data Bank of Japan, the European Molecular Biology Laboratory, and GenBank database under the accession number AB024327. Back

2 To whom requests for reprints should be addressed, at Nagoya University School of Medicine, Department of Mol. Pathogen., 65 Tsurumai-cho, Showa-ku, Nagoya, Japan. Phone: 81-52-744-2463; Fax: 81-52-744-2464; E-mail: smatsuda{at}tsuru.med.nagoya-u.ac.jp Back

3 The abbreviations used are: TGF, transforming growth factor; WD, Trip-Asp; RT-PCR, reverse transcription-PCR; RH, radiation hybrid; GST, glutathione S-transferase; MAPK, mitogen-activated protein kinase; ORF, open reading frame. Back

Received 10/ 4/99. Accepted 11/15/99.


   REFERENCES
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results and Discussion
 REFERENCES
 

  1. Chen R-H., Miettinen P. J., Muraoka E. M., Choy L., Derynck R. A WD-domain protein that is associated with and phosphorylated by the type II TGF-ß receptor. Nature (Lond.), 377: 548-552, 1995.[Medline]
  2. van der Voorn L., Ploegh H. L. The WD-40 repeat. FEBS Lett., 307: 131-134, 1992.[Medline]
  3. Neer E. J., Schmidt C. J., Nambudripad R., Smith T. F. The ancient regulatory-protein family of WD-repeat proteins. Nature (Lond.), 371: 297-300, 1994.[Medline]
  4. Neer E. J., Schmidt C. J. G protein heterodimers: new structures propel new questions. Cell, 84: 175-178, 1996.[Medline]
  5. Lenormand P., Brondello J. M., Brunet A., Pouyssegur J. Growth factor-induced p42/p44 MAPK nuclear translocation and retention requires both MAPK activation and neosynthesis of nuclear anchoring proteins. J. Cell Biol., 10: 625-633, 1998.
  6. Emig R., Magener A., Ehemann V., Meyer A., Stilgenbauer F., Volkmann M., Wallwiener D., Sinn H. P. Aberrant cytoplasmic expression of the p16 protein in breast cancer is associated with accelerated tumour proliferation. Br. J. Cancer, 78: 1661-1668, 1998.[Medline]
  7. Altschul G. F., Gish W., Miller W., Mayers E. W., Lipman D. J. Basic local alignment search tool. J. Mol. Biol., 215: 403-410, 1990.[Medline]
  8. Matsuda S. K., Tsuzuku J., Ohsugi M., Yoshida M., Emi M., Nakamura Y., Onda M., Yoshida Y., Nishiyama A., Yamamoto T. Tob, a novel protein that interacts with p185erbB2, is associated with anti-proliferative activity. Oncogene, 12: 705-713, 1996.[Medline]
  9. Satomi K., Matsuda S., Matsuda Y., Saitoh T., Ohsugi M., Yamamoto T. Molecular cloning and characterization of Byp, a murine receptor-type tyrosine phosphatase similar to human DEP-1. FEBS Lett., 378: 7-14, 1996.[Medline]
  10. Wickens M. How the messenger got its tail: addition of poly(A) in the nucleus. Trends Biochem. Sci., 15: 277-281, 1990.[Medline]
  11. Datta P. K., Chytil A., Gorska A. E., Moses H. L. Identification of STRAP, a novel WD domain protein in transforming growth factor-ß signaling. J. Biol. Chem., 273: 34671-34674, 1998.[Abstract/Free Full Text]
  12. Ueki N., Seki N., Yano K., Masuho Y., Saito T., Muramatsu M. Isolation and characterization of a novel human gene (HFB30) which encodes a protein with a RING finger motif. Biochim. Biophys. Acta, 14: 232-236, 1999.
  13. Takahashi K., Sawasaki Y., Hata J., Mukai K., Goto T. Spontaneous transformation and immortalization of human endothelial cells. In Vitro Cell Dev. Biol., 26: 265-274, 1990.[Medline]
  14. Hamaguchi M., Grandri C., Hanafusa H. Phosphorylation of cellular proteins in Rous sarcoma virus-infected cells: analysis by use of anti-phosphotyrosine antibodies. Mol. Cell. Biol., 8: 3035-3042, 1988.[Abstract/Free Full Text]
  15. Hartsough M. T., Frey R. S., Zipfel P. A., Buard A., Cook S. J., McCormick F., Mulder K. M. Altered transforming growth factor ß signaling in epithelial cells when Ras activation is blocked. J. Biol. Chem., 271: 22368-22375, 1996.[Abstract/Free Full Text]
  16. Axmann A., Seidel D., Reimann T., Hempel U., Wenzel K-W. Transforming growth factor-ß1-induced activation of the Raf-MEK-MAPK signaling pathway in Rat lung fibroblasts via a PKC- dependent mechanism. Biochem. Biophys. Res. Commun., 249: 456-460, 1998.[Medline]
  17. Burgering B. M., de Vries-Smits A. M., Medema R. H., van Weeren P. C., Tertoolen L. G., Bos J. L. Epidermal growth factor induces phosphorylation of extracellular signal-regulated kinase 2 via multiple pathways. Mol. Cell. Biol., 13: 7248-7256, 1993.[Abstract/Free Full Text]
  18. Kuno Y., Kondo K., Iwata H., Senga T., Akiyama S., Ito K., Takagi H., Hamaguchi M. Tumor-specific activation of mitogen-activated protein kinase in human colorectal and gastric carcinoma tissues. Jpn. J. Cancer Res., 89: 903-909, 1998.[Medline]
  19. Watson M. A., Fleming T. P. Mammaglobin, a mammary-specific member of the uteroglobin gene family, is overexpressed in human breast cancer. Cancer Res., 56: 860-865, 1996.[Abstract/Free Full Text]
  20. Emig R., Magener A., Ehemann V., Meyer A., Stilgenbauer F., Volkmann M., Wallwiener D., Sinn H-P. Aberrant cytoplasmic expression of the p16 protein in breast cancer is associated with accelerated tumour proliferation. Br. J. Cancer, 78: 1661-1668, 1998.



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