Clinical Cancer Research CTRC-AACR San Antonio Breast Cancer Symposium
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Perera, A. D.
Right arrow Articles by Walker, C. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Perera, A. D.
Right arrow Articles by Walker, C. L.
Clinical Cancer Research Vol. 6, 1518-1523, April 2000
© 2000 American Association for Cancer Research

Experimental Therapeutics, Preclinical Pharmacology

Requirement for the von Hippel-Lindau Tumor Suppressor Gene for Functional Epidermal Growth Factor Receptor Blockade by Monoclonal Antibody C225 in Renal Cell Carcinoma1

Ayesh D. Perera, Elena V. Kleymenova and Cheryl L. Walker2

The University of Texas M. D. Anderson Cancer Center, Science Park Research Division, Smithville, Texas 78957


   ABSTRACT
Top
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Renal cell carcinoma (RCC) is a cytologically and histologically diverse disease in which a spectrum of distinct molecular alterations occurs, including the inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene, which is specific for the clear cell variant of RCC. The prognosis for RCC is poor, and, to date, no effective systemic treatment is available for this cancer. In the present study, we assessed the extent to which the transforming growth factor {alpha}-epidermal growth factor receptor (EGFR) autocrine loop could be used as a potential therapeutic target for RCC. Northern blot analysis of transforming growth factor {alpha} and EGFR revealed variable but consistent expression of these transcripts in cell lines derived from both clear cell and non-clear cell RCC variants, indicating the potential for this autocrine loop in both tumor types. The therapeutic utility of interruption of this feedback loop was determined by examining growth inhibition after the exposure of these cell lines to a humanized anti-EGFR monoclonal antibody, C225. In vitro treatment of clear cell RCC-derived cell lines lacking VHLresulted in only a modest decrease in growth rate. In contrast, non-clear cell RCC-derived cell lines that retained VHL responded significantly to C225 treatment. Transfection of VHL into VHL-negative RCC cell lines restored responsiveness to C225, indicating that this tumor suppressor gene is required for effective EGFR blockade. Growth inhibition by C225 in VHL-positive cells was linked to a requirement for VHL to up-regulate p27 in response to C225. These data provide compelling evidence that treatment modalities for RCC are likely to be strongly influenced by the molecular etiology of this phenotypically diverse cancer.


   INTRODUCTION
Top
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
RCC3 is a phenotypically heterogeneous neoplasm whose early detection is hindered by the absence of early clinical manifestations. Subsequent therapy of RCC is impaired by both a high incidence of metastasis at the time of presentation (1) and resistance to conventional cytotoxic treatment regimens (2 , 3) . Increased presentation of this cancer (a 38% increase was observed between 1974 and 1990) is thought to be linked to an increase in smoking, obesity, high-protein diet, and hypertension and, to a lesser degree, to an earlier diagnosis (4) . Diverse histological and cytological variants are a hallmark of RCC, and distinct molecular etiologies are thought to underlie the different phenotypes observed in this disease. The most common type of RCC, solid RCC of the clear cell type, is thought to arise as a result of loss of function of the VHL tumor suppressor gene (5, 6, 7) . Other RCC variants, which can be collectively classified as non-clear cell RCC, have various genetic alterations (8, 9, 10) but no VHL defects (6 , 7) . In light of these differences, it is not unreasonable to propose that the outcome of treatment modalities for this disease may be strongly influenced by the molecular etiology of a particular RCC variant. The recent demonstration in a retrospective study that effectiveness of RCC treatment with cytokines correlated with tumor histology (11) provides support for this hypothesis.

In RCC, as in many other cancers, the TGF-{alpha}-EGFR autocrine loop is regulated aberrantly as compared to epithelial cells of the proximal tubule of the renal cortex, from which RCC originates (12, 13, 14) . TGF-{alpha} is a potent mitogen, and its interaction with the EGFR is thought to play a significant role in neoplastic transformation and the subsequent deregulated cell growth observed during tumorigenesis. Interruption of the TGF-{alpha}-EGFR loop may therefore provide a potential therapeutic for the treatment of RCC. In the present study, we explored this possibility by using a humanized monoclonal antibody against the EGFR, C225, on cell lines derived from both clear and non-clear cell RCC. This antibody has been shown to be effective in suppressing the growth and, in some cases, inducing apoptosis (15) in cancer cell lines of epithelioid origin (16, 17, 18) and in RCC xenografts in nude mice (16 , 19) . In the present study, we report the anti-proliferative effect of C225 on RCC-derived cell lines and provide evidence that the molecular etiology of the RCC variant may have a strong influence on the efficacy of this treatment. Specifically, RCC variants retaining VHL exhibited a significantly greater growth inhibition upon C225 treatment as compared with RCC cells that harbor VHL mutations. Responsiveness of VHL-positive RCC cells to C225 appears to be linked to the ability of these cells to up-regulate the cyclin-dependent kinase inhibitor p27 in response to EGFR blockade.


   MATERIALS AND METHODS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Reagents.
The humanized anti-EGFR monoclonal antibody C225 has been described previously (20 , 21) and was kindly provided by Imclone Systems (New York, NY). The p27 monoclonal antibody used for Western blot analysis was obtained from Transduction Laboratories (Lexington, KY).

Tumor Cell Lines and Culture Conditions.
A panel of cell lines derived from clear cell and non-clear cell RCCs that have previously been characterized for VHL mutations (22) was used. Cell lines 112, 124, 786-0, and 101 and their derivatives stably transfected with either wild-type or mutant VHL constructs (23) were a gift from Dr. J. Gnarra (Louisiana State University Medical Center, New Orleans, LA). Cell line 786-0/g7f expresses a FLAG-tagged full-length VHL protein (tagged at the COOH terminus), and cell line 786-0/157{Delta} expresses VHL amino acids 1–157, with amino acids 158–213 being deleted. Cell line 101wt expresses the full-length VHL protein. Cell lines 112 and 124 were derived from papillary tumors and have been previously shown to contain wild-type VHL (22) . Cell lines ACHN, CAKI-1, and A498 were purchased from the American Type Culture Collection (Manassas, VA). Nontransfected cells were cultured in DMEM or MEM containing 10% FBS. Cell lines transfected with either wild-type or mutant VHL were grown in DMEM with 10% FBS and 800 µg/ml G418. To confirm VHL status in the transfected cell lines used in the study, total RNA was obtained by conventional CsCl centrifugation, and reverse transcription-PCR was performed with a forward primer located in exon 1 (222–5'-CATCTTCTGCAATCGCAGTCCGCGCGT- 3') and a reverse primer located in the vector (5'-ACTTTCCACACCCTAACTGACACACAT- 3'). As expected, PCR using primers located in exon 1 and the vector did not result in VHL cDNA amplification in cell line 101. In contrast, cell lines bearing VHL constructs had reverse transcription-PCR products of the expected size of 501 bp for the wild-type constructs and 333 bp for the mutant VHL constructs (data not shown).

RNA Isolation and Northern Analysis.
Total RNA was isolated from cells in log phase by conventional centrifugation in CsCl, and poly(A)+ RNA was obtained by affinity chromatography. Five µg of poly(A)+ RNA from each cell line were used for Northern blot analysis using the standard protocol. Blots were hybridized with 32P-labeled probes for TGF-{alpha} (24) and EGFR (25) .

Proliferation Assays.
Four RCC cell lines with mutant VHL (VHL-) and four RCC cell lines with wild-type VHL (VHL+) were plated in triplicate into 24-well dishes. Twenty-four h before exposure to C225, cells were switched to low-serum growth conditions (0.5% FBS in growth medium). Cells were treated with 80 or 160 nM C225 or vehicle, and the number of cells in each well at different time points was determined using a Coulter counter. For some cell lines, the experiment was repeated three times. Statistical analysis was performed using StatView (Abacus Concepts, Berkeley, CA). Percentage of inhibition at each time point was determined as a ratio of a mean number of cells treated with C225:vehicle-treated cells.

Analysis of Cell Cycle and p27 Expression in Response to C225.
Isogenic cell lines 786-0/157{Delta} and 786-0/g7f were grown in media containing 10% FBS. When cells were at 20–40% confluence, they were switched to the low-serum conditions (0.5% FBS in growth media) for 24 h. Cells were then exposed to 160 nM C225 and harvested at 24 and 48 h for either protein or DNA content analysis. For protein analysis, total cell lysates were obtained by resuspending cells in PBS containing 0.1% SDS, 0.01% NP40, 4 mg/ml aprotinin, 0.1 M phenylmethylsulfonyl fluoride, 25 mg/ml soybean trypsin inhibitor, and 50 mM leupeptin. Cell lysates with an equal amount of total protein were electrophoresed on 4–20% Tris-glycine gradient gels (Novex, San Diego, CA) and transferred to polyvinylidene difluoride membranes (Pierce, Rockford, IL). Blots were probed with a p27 monoclonal antibody, and a protein detector Western blot kit, LumiGLO System (Kirkegaard and Perry Laboratories, Gaithersburg, MD), was used to visualize the bands. For DNA content analysis, cells were trypsinized, washed twice in ice-cold PBS, and resuspended in 200 µl of PBS. Cells were fixed by slowly adding the cell suspension drop-wise to 4 ml of ice-cold 70% ethanol with gentle agitation and incubated on ice for 30 min. The suspension was then centrifuged, and cells were resuspended in 1 ml of PBS containing 40 µg/ml propidium iodide and 100 µg/ml RNase A and incubated at 37°C for 30 min. DNA content analysis was performed on a Coulter EPICS Elite flow cytometer, and data were analyzed using MultiCycle DNA cell cycle analysis software (Phoenix Flow Systems, San Diego, CA). Aggregate discrimination mode was used to analyze single cells only.


   RESULTS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Expression of TGF-{alpha} and EGFR mRNA.
A panel of RCC cell lines derived from clear cell (mutant VHL) and non-clear cell (wild-type VHL) tumors was examined for expression of mRNA transcripts for TGF-{alpha} and EGFR to establish the potential for blockade of this autocrine loop in different RCC variants. The three expected RNA transcripts of EGFR were detected in all cell lines, with the 10.3-kb transcript being the most abundant (Fig. 1)Citation . Although expression was variable, TGF-{alpha} RNA transcripts were also detected in all of the cell lines examined, indicating that the TGF-{alpha}-EGFR autocrine loop was present in both VHL+ and VHL- RCC cell lines and was not a hallmark of any specific histological variant.



View larger version (59K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 1. Northern blot analysis of EGFR and TGF-{alpha} RNA expression in RCC cell lines. A major 10.3-kb EGFR transcript was detected in both clear cell (VHL-) and non-clear cell (VHL+) RCC cell lines. The slight difference in the band size is an artifact resulting from differences in electrophoretic mobility in different experiments. Normalization to glyceraldehyde-3-phosphate dehydrogenase revealed variable expression of both EGFR and TGF-{alpha} in different RCC histological variants.

 
Effect of C225 on Cell Proliferation.
The panel of RCC-derived cell lines, including VHL- and VHL+ isogenic cell lines, was examined for growth inhibition by C225. During log phase, maximum growth inhibition of all cell lines was observed at day 4 after C225 treatment. A summary of growth inhibition at this time point for all cell lines is shown in Table 1Citation . Statistical analysis of inhibition data revealed that the antiproliferative effect of C225 on VHL- cell lines was, in most instances, only marginal, with an average maximal growth suppression of 14.58 ± 6.22%. In contrast, cell lines with wild-type VHL had, on average, a 42.25 ± 1.88% inhibition of growth, which was significantly greater by unpaired Student’s test (P < 0.02) compared with cell lines with mutant VHL.


View this table:
[in this window]
[in a new window]

  		Table 1 Maximum inhibition of RCC cell lines by C225

 
The specific contribution of VHL in mediating the effect of the C225 antibody on cell growth was demonstrated in two clear cell RCC-derived cell lines, 786-0 and 101, which have mutant VHL. In both cell lines, the introduction of wild-type VHL dramatically changed the profile of growth inhibition, conferring responsiveness to C225 on VHL- cells (Fig. 2)Citation . Interestingly, a statistically significant growth inhibition effect of C225 became evident much earlier in VHL+ cells (days 3 and 2 for 101wt and 786-0/g7f, respectively) than in VHL- cells (day 5 for both 101 and 786-0/157{Delta}).



View larger version (26K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 2. VHL impact on growth kinetics of RCC cell lines treated with C225. VHL- cell lines 101 and 786-0/157{Delta} and isogenic matched VHL+ cell lines 101wt and 786-0/g7f were treated with 160 nM C225, and growth kinetics were obtained. {circ} and •, mean number of untreated and treated cells, respectively, determined for three wells at each time point.

 
Effect of C225 on Cell Cycle and p27 Expression.
To determine whether inhibition of growth of VHL+ cells by C225 affects the progression of the cell cycle, DNA content was determined in 786-0/g7f cells at various time points after the treatment. After 24 h of exposure to this antibody, DNA content analysis demonstrated that the percentage of treated cells in S phase had decreased from 29.5% to 13.8%, with a commensurate increase in the G1 fraction from 68% to 85%, suggesting that C225 treatment had induced a G1 arrest in these cells (Fig. 3)Citation . The same distribution of cells within the cell cycle phases was observed at 48 and 96 h after the treatment. Interestingly, the sub-2N fraction of cells also increased in the C225-treated cells, suggesting a possible induction of apoptosis in RCC by C225 (data not shown).



View larger version (27K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 3. Effect of C225 treatment on the cell cycle. Exposure of the VHL+ RCC cell line 786-0/g7f to 160 nM C225 decreased the number of cells in G2 and S phases and G1 cell cycle arrest.

 
The VHL gene product has been previously shown to regulate response to growth factor deprivation, and this activity has been attributed to the ability of VHL to regulate p27 levels (26) . Up-regulation of p27 in tumors other than RCC in response to C225 has been previously demonstrated (17 , 18) ; therefore, we examined the involvement of p27 in growth inhibition by C225 in RCC. Western blot analysis of p27 revealed that exposure of the VHL- cell line 786-0/157{Delta} to C225 did not change the level of expression of this protein (Fig. 4)Citation . In contrast, treatment of the isogenic matched cell line 786-0/g7f (transfected with the wild-type VHL) with C225 increased the p27 level after 24 h of exposure (Fig. 4)Citation .



View larger version (17K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 4. Up-regulation of p27 levels in VHL+ RCC cells in response to C225. Levels of p27 were measured by Western blot analysis in VHL+ (wt VHL) and VHL- (mutant VHL) derivatives of the 786-0 RCC cell line grown in media with 160 nM C225 (AB) or without (-) C225 after 24 and 48 h of treatment.

 

   DISCUSSION
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
The relative ineffectiveness of traditional systemic chemotherapeutic modalities for RCC clearly warrants the exploration of novel approaches to the treatment of this disease. In the present study, we report the effectiveness of the humanized anti-EGFR monoclonal antibody C225 in suppressing the growth of a panel RCC cell lines derived from clear cell and non-clear cell tumors. Although the growth-inhibitory action of C225 on RCC cell lines has been reported previously (15 , 16 , 27) , possible differences in response between RCC tumor variants to this potential therapeutic agent have not been explored. Consistent with the previously reported TGF-{alpha}-EGFR autocrine loop in RCC (13 , 27 , 28) , our Northern blot analysis detected the expression of both of EGFR and TGF-{alpha} RNA transcripts in cell lines derived from different RCC histological variants, namely, clear and non-clear cell RCCs. We then investigated whether the response to C225 was affected by the molecular etiology of the RCC cell lines and hence the tumor type. The exposure of a panel of cell lines to C225 revealed a distinct dichotomy in response to this agent. Non-clear cell RCC cell lines with the wild-type VHL responded robustly to C225, whereas the cell lines that originated from clear cell variants of RCC and had mutations in the VHL gene showed a generally weaker response to this antibody. How the level of expression of either TGF-{alpha} or EGFR influences the response of RCC cells to C225 is not clear at present and requires further investigation. However, the TGF-{alpha}-EGFR autocrine loop appears to be in place in both histological RCC variants studied, suggesting that, in addition to the autocrine loop, other molecular events impact the C225 inhibitory effect on RCC.

RCC of the clear cell variant is generally thought to result from VHL inactivation (5, 6, 7) , whereas this gene is unaltered in non-clear cell variants (6 , 7) . The different response observed in VHL- and VHL+ RCC variants in the present study suggested that the VHL gene in RCC could mediate the inhibitory effects of C225. Direct evidence that the presence of wild-type VHL accounted for this difference was obtained for clear cell RCC cell lines 101 and 786-0, which had been shown to be poor responders to C225 (Table 1)Citation . These cell lines, after transfection with wild-type VHL, have shown a dramatic transition from a relatively weak response to a robust response to C225 treatment (Table 1Citation ; Fig. 2Citation ). Thus our data indicate that VHL function may be required for the therapeutic action of C225 in the treatment of RCC.

How VHL mediates the downstream events of EGFR blockade by C225 is not clear at present. The function or probable functions of VHL are currently being elucidated (29) . To date, the protein product of this tumor suppressor gene is thought to be involved in a number of pathways including interaction with the elongin BC complex (30 , 31) , protein degradation (32) , and negative regulation of hypoxia-inducible genes such as VEGF via interaction with Sp1 (33) or via posttranscriptional mechanisms (23) . In the context of the present experiments, the role of VHL interaction with elongin BC in protein degradation is of special interest. Pause et al. (26) demonstrated that VHL function was necessary for RCC cell line 786-0 to up-regulate p27 and exit the cell cycle in response to serum starvation. This, as suggested by the authors, may result from the inability of mutant VHL to inhibit Hs-Cul-2-mediated p27 degradation. Consistent with this hypothesis, we found that the extent of C225 inhibition of cell growth and p27 up-regulation in RCC is significantly dependent on VHL function. The relative lack of response of the clear cell-derived RCC cell lines to C225 appears to be due to their inability to increase p27 levels. Our findings that VHL- RCC cell lines were unable to augment p27 in response to C225 and that the reintroduction of wild-type VHL can overcome this defect clearly point to a requirement for VHL function in p27 up-regulation and growth inhibition in response to C225. Interestingly, it has been shown for other tumors, which are not associated with alterations at the VHL locus, that the inhibition of cell growth by C225 is also mediated by the up-regulation of p27 (17 , 34) .

In summary, the present experiments show that blockade of the EGFR by the monoclonal antibody C225 may provide a potential therapeutic avenue for the treatment of RCC. However, the dramatic difference in response between different RCC variants clearly indicates that the molecular etiology of these tumors may have a critical impact on the utility of this and possibly other potential therapeutic agents in the treatment of this disease.


   ACKNOWLEDGMENTS
 
We thank Drs. John Mendelsohn, James Gnarra, and Zhen Fan for helpful discussions and critical review of the manuscript; Imclone, Inc. for donations of C225; and Ken Claypool for assistance with fluorescence-activated cell-sorting analysis.


   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 Supported in part by NIH Grant CA63613 and National Institute of Environmental Health Sciences Center Grant ES07784 Back

2 To whom requests for reprints should be addressed, at The University of Texas M. D. Anderson Cancer Center, Science Park Research Division, Box 389, Smithville, TX 78957. Phone: (512) 237-9525; Fax: (512) 237-2475. Back

3 The abbreviations used are: RCC, renal cell carcinoma; EGFR, epidermal growth factor receptor; TGF-{alpha}, transforming growth factor {alpha}; VHL, von Hippel-Lindau; FBS, fetal bovine serum. Back

Received 9/ 7/99; revised 1/ 5/00; accepted 1/11/00.


   REFERENCES
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. DeKiernion, J. B., and Belldegrun, A. Renal tumors. In: M. F. Campbell, A. B. Retik, E. Vaughn, and P. C. Walsh (eds.), Campbell’s Urology, 6th ed., pp. 1053–1093. Philadelphia, 1993.
  2. Finney R. The value of radiotherapy in the treatment of hypernephroma: a clinical trial. Br. J. Urol., 45: 258-269, 1973.[Medline]
  3. Yagoda A., Abi-Rached B., Petrylak D. Chemotherapy for advanced renal-cell carcinoma: 1983–1993. Semin. Oncol., 22: 42-60, 1995.[Medline]
  4. Figlin R. A. Renal cell carcinoma: management of advanced disease. J. Urol., 161: 381-386, 1999.[CrossRef][Medline]
  5. Latif F., Tory K., Gnarra J., Yao M., Duh F. M., Orcutt M. L., Stackhouse T., Kuzmin I., Modi W., Geil L., Schmidt L., Zhou F., Li M., Wei M. H., Chen F., Glenn G., Choyke P., Walther M. M., Weng Y., Duar D. R., Dean M., Glavac D., Richards F. M., Crossey P. A., Ferguson-Smith M. A., Paslter D. L., Chumakov I., Cohen D., Chinault A. G., Maher E. R., Linehan W. M., Zbar B., Lerman M. I. Identification of the von Hippel-Lindau disease tumor suppressor gene. Science (Washington DC), 260: 1317-1320, 1993.[Abstract/Free Full Text]
  6. Kenck C., Wilhelm M., Bugert P., Staehler G., Kovacs G. Mutation of the VHL gene is associated exclusively with the development of non-papillary renal cell carcinomas. J. Pathol., 179: 157-161, 1996.[CrossRef][Medline]
  7. Foster K., Prowse A., van den Berg A., Fleming S., Hulsbeek M. M., Crossey P. A., Richards F. M., Cairns P., Affara N. A., Ferguson-Smith M. A., Buys C. H. M., Maher E. R. Somatic mutations of the von Hippel-Lindau disease tumor suppressor gene in non-familial clear cell renal carcinoma. Hum. Mol. Genet., 3: 2169-2173, 1994.[Abstract/Free Full Text]
  8. Jiang F., Richter J., Schraml P., Bubendorf L., Gasser T., Sauter G., Mihatsch M. J., Moch H. Chromosomal imbalances in papillary renal cell carcinoma. Am. J. Pathol., 153: 1467-1473, 1998.[Abstract/Free Full Text]
  9. Clark J., Lu Y-J., Sidhar S. K., Parker C., Gill S., Smedley D., Hamoudi R., Linehan W. M., Shipley J., Cooper C. Fusion of splicing factor genes PSF and NonO(p54nrb) to the TFE3 gene in papillary renal cell carcinoma. Oncogene, 15: 2233-2239, 1997.[CrossRef][Medline]
  10. Schwerdtle R. F., Storkel S., Neuhaus C., Brauch H., Weidt E., Brenner W., Hohenfellner R., Huber C., Decker H-J. Allelic losses at chromosomes 1p, 2p, 6p, 10p, 13q, 17p, and 21q significantly correlate with the chromophobe subtype of renal cell carcinoma. Cancer Res., 56: 2927-2930, 1996.[Abstract/Free Full Text]
  11. Wu J., Caliendo G., Hu X. P., Dutcher J. P. Impact of histology on the treatment outcome of metastatic or recurrent renal cell carcinoma. Med. Oncol., 15: 44-49, 1998.[Medline]
  12. Freeman M. R., Washecka R., Chung L. W. Aberrant expression of epidermal growth factor receptor and HER-2 (erbB-2) messenger RNAs in human renal cancers. Cancer Res., 49: 6221-6225, 1989.[Abstract/Free Full Text]
  13. Mydlo J. H., Michaeli J., Cordon-Cardo C., Goldenberg A. S., Heston W. D., Fair W. R. Expression of transforming growth factor {alpha} and epidermal growth factor receptor messenger RNA in neoplastic and nonneoplastic human kidney tissue. Cancer Res., 49: 3407-3411, 1989.[Abstract/Free Full Text]
  14. Salomon D. S., Brandt R., Ciardiello F., Normanno N. Epidermal growth factor-related peptides and their receptors in human malignancies. Crit. Rev. Oncol. Hematol., 19: 183-232, 1995.[Medline]
  15. Ciardiello F., Caputo R., Bianco R., Damiano V., Pomatico G., Pepe S., Bianco A. R., Agrawal S., Mendelsohn J., Tortora G. Cooperative inhibition of renal cancer growth by anti-epidermal growth factor receptor antibody and protein kinase A antisense oligonucleotide. J. Natl. Cancer Inst., 90: 1087-1094, 1998.[Abstract/Free Full Text]
  16. Perrotte P., Matsumoto T., Inoue K., Kuniyasu H., Eve B. Y., Hicklin D. J., Radinsky R., Dinney C. P. Anti-epidermal growth factor receptor antibody C225 inhibits angiogenesis in human transitional cell carcinoma growing orthotopically in nude mice. Clin. Cancer Res., 5: 257-265, 1999.[Abstract/Free Full Text]
  17. Fan Z., Shang B. Y., Lu Y., Chou J. L., Mendelsohn J. Reciprocal changes in p27Kip1 and p21Cip1 in growth inhibition mediated by blockade or overstimulation of epidermal growth factor receptors. Clin. Cancer Res., 3: 1943-1948, 1997.[Abstract]
  18. Peng D., Fan Z., Lu Y., DeBlasio T., Scher H., Mendelsohn J. Anti-epidermal growth factor receptor monoclonal antibody 225 up-regulates p27KIP1 and induces G1 arrest in prostatic cancer cell line DU145. Cancer Res., 56: 3666-3669, 1996.[Abstract/Free Full Text]
  19. Prewett M., Rothman M., Waksal H., Feldman M., Bander N. H., Hicklin D. J. Mouse-human chimeric anti-epidermal growth factor receptor antibody C225 inhibits the growth of human renal cell carcinoma xenografts in nude mice. Clin. Cancer Res., 4: 2957-2966, 1998.[Abstract]
  20. Kawamoto T., Sato J. D., Le A., Polikoff J., Sato G. H., Mendelsohn J. Growth stimulation of A431 cells by epidermal growth factor: identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Proc. Natl. Acad. Sci. USA, 80: 1337-1341, 1983.[Abstract/Free Full Text]
  21. Goldstein N. I., Prewett M., Zuklys K., Rockwell P., Mendelsohn J. Biological efficacy of a chimeric antibody to the epidermal growth factor receptor in a human tumor xenograft model. Clin. Cancer Res., 1: 1311-1318, 1995.[Abstract]
  22. Gnarra J. R., Tory K., Weng Y., Schmidt L., Wei M. H., Li H., Latif F., Liu S., Chen F., Duh F. M., Lubensky L., Duar D. R., Florence C., Pozzatti R., Walther M. M., Bander N. H., Grossman H. B., Brauch M., Pomer S., Brooks J. D., Isaacs W. B., Lerman M. I., Zbar B., Linehan W. M. Mutations of the VHL tumour suppressor gene in renal carcinoma. Nat. Genet., 7: 85-90, 1994.[CrossRef][Medline]
  23. Gnarra J. R., Zhou S., Merrill M. J., Wagner J. R., Krumm A., Papavassiliou E., Oldfield E. H., Klausner R. D., Linehan W. M. Post-transcriptional regulation of vascular endothelial growth factor mRNA by the product of the VHL tumor suppressor gene. Proc. Natl. Acad. Sci. USA, 93: 10589-10594, 1996.[Abstract/Free Full Text]
  24. Derynck R., Roberts A. B., Winkler M. E., Chen E. Y., Goeddel D. V. Human transforming growth factor-{alpha}: precursor structure and expression in E. coli. Cell, 38: 287-297, 1984.[CrossRef][Medline]
  25. Simmen F. A., Gope M. L., Schulz T. Z., Wright D. A., Carpenter G., O’Malley B. W. Isolation of an evolutionarily conserved epidermal growth factor receptor cDNA from human A431 carcinoma cells. Biochem. Biophys. Res. Commun., 124: 125-132, 1984.[Medline]
  26. Pause A., Lee S., Lonergan K. M., Klausner R. D. The von Hippel-Lindau tumor suppressor gene is required for cell cycle exit upon serum withdrawal. Proc. Natl. Acad. Sci. USA, 95: 993-998, 1998.[Abstract/Free Full Text]
  27. Atlas I., Mendelsohn J., Baselga J., Fair W. R., Masui H., Kumar R. Growth regulation of human renal carcinoma cells: role of transforming growth factor {alpha}. Cancer Res., 52: 3335-3339, 1992.[Abstract/Free Full Text]
  28. Ramp U., Jaquet K., Reinecke P., Schardt C., Friebe U., Nitsch T., Marx N., Gabbert H. E., Gerharz C. D. Functional intactness of stimulatory and inhibitory autocrine loops in human renal carcinoma cell lines of the clear cell type. J. Urol., 157: 2345-2350, 1997.[CrossRef][Medline]
  29. Kaelin W. G., Iliopoulos O., Lonergan K. M., Ohh M. Functions of the von Hippel-Lindau tumour suppressor protein. J. Intern. Med., 243: 535-539, 1998.[CrossRef][Medline]
  30. Duan D. R., Pause A., Burgess W. H., Aso T., Chen D. Y., Garrett K. P., Conaway R. C., Conaway J. W., Linehan W. M., Klausner R. D. Inhibition of transcription elongation by the VHL tumor suppressor protein. Science (Washington DC), 269: 1402-1406, 1995.[Abstract/Free Full Text]
  31. Kibel A., Iliopoulos O., DeCaprio J. A., Kaelin W. G., Jr. Binding of the von Hippel-Lindau tumor suppressor protein to elongin B and C. Science (Washington DC), 269: 1444-1446, 1995.[Abstract/Free Full Text]
  32. Kamura T., Sato S., Haque D., Liu L., Kaelin W. G., Jr., Conaway R. C., Conaway J. W. The elongin BC complex interacts with the conserved SOCS-box motif present in members of the SOCS, ras, WD-40 repeat, and ankyrin repeat families. Genes Dev., 12: 3872-3881, 1998.[Abstract/Free Full Text]
  33. Mukhopadhyay D., Knebelmann B., Cohen H. T., Ananth S., Sukhatme V. P. The von Hippel-Lindau tumor suppressor gene product interacts with Sp1 to repress vascular endothelial growth factor promoter activity. Mol. Cell. Biol., 17: 5629-5639, 1997.[Abstract]
  34. Wu X., Rubin M., Fan Z., DeBlasio T., Soos T., Koff A., Mendelsohn J. Involvement of p27KIP1 in G1 arrest mediated by an anti-epidermal growth factor receptor monoclonal antibody. Oncogene, 12: 1397-1403, 1996.[Medline]



This article has been cited by other articles:


Home page
 Clin. Cancer Res.Home page
T. G. Johns, R. M. Perera, S. C. Vernes, A. A. Vitali, D. X. Cao, W. K. Cavenee, A. M. Scott, and F. B. Furnari
The Efficacy of Epidermal Growth Factor Receptor-Specific Antibodies against Glioma Xenografts Is Influenced by Receptor Levels, Activation Status, and Heterodimerization
Clin. Cancer Res., March 15, 2007; 13(6): 1911 - 1925.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
A. Franovic, I. Robert, K. Smith, G. Kurban, A. Pause, L. Gunaratnam, and S. Lee
Multiple Acquired Renal Carcinoma Tumor Capabilities Abolished upon Silencing of ADAM17
Cancer Res., August 15, 2006; 66(16): 8083 - 8090.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
A. Potti and D. J. George
Tyrosine Kinase Inhibitors in Renal Cell Carcinoma
Clin. Cancer Res., September 15, 2004; 10(18): 6371S - 6376S.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
J. E. Dancey
Epidermal Growth Factor Receptor and Epidermal Growth Factor Receptor Therapies in Renal Cell Carcinoma: Do We Need a Better Mouse Trap?
J. Clin. Oncol., August 1, 2004; 22(15): 2975 - 2977.
[Full Text] [PDF]

Home page
 Cancer Res.Home page
M. Y. Liu, L. Poellinger, and C. L. Walker
Up-Regulation of Hypoxia-inducible Factor 2{alpha} in Renal Cell Carcinoma Associated with Loss of Tsc-2 Tumor Suppressor Gene
Cancer Res., May 15, 2003; 63(10): 2675 - 2680.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
N. de Paulsen, A. Brychzy, M.-C. Fournier, R. D. Klausner, J. R. Gnarra, A. Pause, and S. Lee
Role of transforming growth factor-alpha in von Hippel- Lindau (VHL)-/- clear cell renal carcinoma cell proliferation: A possible mechanism coupling VHL tumor suppressor inactivation and tumorigenesis
PNAS, February 1, 2001; (2001) 31587498.
[Abstract] [Full Text]

Home page
 Proc. Natl. Acad. Sci. USAHome page
N. de Paulsen, A. Brychzy, M.-C. Fournier, R. D. Klausner, J. R. Gnarra, A. Pause, and S. Lee
Role of transforming growth factor-alpha in von Hippel- Lindau (VHL)-/- clear cell renal carcinoma cell proliferation: A possible mechanism coupling VHL tumor suppressor inactivation and tumorigenesis
PNAS, February 13, 2001; 98(4): 1387 - 1392.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Perera, A. D.
Right arrow Articles by Walker, C. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Perera, A. D.
Right arrow Articles by Walker, C. L.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online