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Departments of 1 Pathology, 2 Hematopathology, 3 Molecular Therapeutics, and 4 Experimental Therapeutics, The University of Texas M. D. Anderson Cancer Center, Houston, Texas; 5 Department of Obstetrics and Gynecology, Research Division, University of British Columbia, Vancouver, British Columbia, Canada; and 6 Department of Medical Oncology, Dana-Farber Cancer Institute and Department of Medicine, Harvard Medical School, Boston, Massachusetts
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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B pathway, which is known to contribute to the tumor growth of naturally ovarian cancer cells. Incubation with antibodies to IL-1ß or IL-8 led to apoptosis in the ras-transformed cells and ovarian cancer cells but not in immortalized cells that had not been transformed. Thus, the transformed human ovarian surface epithelial cells recapitulated many features of natural ovarian cancer including a subtype of ovarian cancer histology, formation of ascites, CA125 expression, and nuclear factor-B-mediated cytokine activation. These cells provide a novel model system to study human ovarian cancer. |
INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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The RAS gene family has been implicated in the development of many human epithelial cancers. RAS genes encode highly homologous and evolutionarily conserved Mr 21,000 GTP-binding proteins, which are often activated in human ovarian cancer. KRAS mutations have been found in 30% of borderline ovarian tumors and in a subset of invasive mucinous ovarian cancers (10, 11, 12, 13, 14, 15)
. In one study, mutations in KRAS or its downstream mediator BRAF were detected in 68% of low-grade serous carcinomas and in 61% of borderline serous tumors, implicating the KRAS pathway in these tumors as well (16)
. The oncogenic allele HRASV12 is present in 
6% of ovarian cancers (15)
. However, physiologically activated HRAS protein is commonly detected in human ovarian cancer, presumably because of an increase in upstream signals from tyrosine kinase growth factor receptors such as Her-2/neu, despite an absence of RAS mutation (17
, 18) . The mechanisms by which ras oncogenes transform human epithelial cells are not clear.
Several investigators have cultured ovarian surface epithelial cells to use them as model system to study ovarian cancer development. Ovarian surface epithelial cells have been cultured from rats (19 , 20) , mice (21 , 22) , and rabbits (23) . A high proportion of rat ovarian surface epithelial cell cultures become spontaneously immortalized and are highly susceptible to K-ras transformation (19) . These spontaneously immortalized cells remained nontumorigenic even after introduction of the SV40 early region expressing large T and small t (T/t) antigens, but they became tumorigenic after transfection with HRAS (24) . Rat ovarian surface epithelial cell lines can also be transformed into tumorigenic cell lines through multiple passages in vitro (20 , 25) . In another approach, Orsulic et al. (26) used an avian retrovirus to introduce c-Myc, K-ras, or Akt into murine ovarian surface epithelial cells and found that introduction two of these oncogenes into cells with a mutated p53 led to the formation of murine ovarian tumors that were similar to human ovarian carcinomas. A more recent study showed that about half of female transgenic mice expressing the transforming region of SV40 under the control of the Mullerian inhibitory substance type II receptor gene promoter developed bilateral ovarian tumors (27) .
The biological differences between human cells and rodent cells make extrapolating results from mice or rats to humans difficult. Ovarian surface epithelial cells from humans are much more difficult to transform than are such cells from rodents. Several laboratories have used cultured human ovarian surface epithelial cells to study human ovarian carcinogenesis (28, 29, 30, 31) . Transfection of such cells with the SV40 T/t or the human papillomavirus type 16 E6/E7 region (29 , 32) can extend their life span, but the transfected cells eventually enter crisis and die. After several months, immortal cells occasionally emerge but do not form colonies in soft agar or tumors in nude mice (29 , 32) . Very rarely, cells transfected with T/t or E6/E7, after many passages in tissue culture, can produce colonies in soft agar or form tumors in nude mice (29 , 30) , presumably as a result of spontaneous mutation. Recently, several nonovarian primary human epithelial and fibroblast cells have been successfully transformed by using a combination of the SV40 T/t, hTERT, and HRASV12 (33 , 34) . Whether a similar combination of genes is sufficient to transform human ovarian surface epithelial cells has not previously been addressed. In addition, the changes in the gene expression profile and the underlying molecular mechanisms associated with ras-mediated transformation in human cells have not been reported. The purpose of this study was to use specific genetic elements to disrupt the signaling pathways implicated in tumorigenesis in human ovarian surface epithelial cells, to assess the molecular mechanisms used by the ras oncoprotein at the genomic level, and to propose a genetic model of the pathophysiology of human ovarian cancer.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Telomerase Assay.
Telomerase activity was measured with the TRAPeze telomerase detection kit (Oncor, Inc., Gaithersburg, MD) as originally described by Kim and Wu (38)
.
Clonogenicity Assay.
Cells (105) were suspended in 2 ml of ovarian epithelial cell medium with 0.35% agarose (Life Technologies, Inc. Rockville, MD), and the suspension was placed on top of 5 ml of solidified 0.7% agarose. Triplicate cultures for each cell type were maintained for 14 days at 37°C on atmosphere of 5% CO2 and 95% air, and fresh medium was added after 1 week. Colonies > 50 µm in diameter were counted after 2 weeks. These experiments were repeated twice.
Tumor Formation in nu/nu Mice.
Equal numbers (5 x 106) of IOSE-80, T80, and T80H cells (or IOSE-29, T29, and T29H cells) were harvested by trypsinization, washed twice with 1x PBS, resuspended in 0.1 ml of saline, and injected either s.c or i.p. into 4–6-week-old BALB/c athymic nude mice (The Jackson Laboratory, Bar Harbor, ME). SKOV-3 ovarian cancer cells were used as a positive control. The mice were kept in a pathogen-free environment and checked every 2 days for 5 months. Mice given i.p. injections were observed for lethargy, poor appetite, and abdominal enlargement. The date at which the first grossly visible tumor appeared was recorded, and tumor size was measured every 2 days thereafter. Mice were killed when tumors reached 1.5 cm in diameter. Tumors were removed and fixed in 10% formalin overnight and subjected to routine histological examination.
Immunohistochemical Analysis.
Tissue sections were deparaffinized and dehydrated, intrinsic peroxidase activity was blocked with a 3% solution of hydrogen peroxide in methanol, and immunostaining was performed with an automated immunohistochemical staining machine (Dako, Carpinteria, CA) and an avidin-biotin-peroxidase enzyme complex using antibodies specific for p53 (1:100 dilution; Dako), WT-1 (1:40 dilution; Dako), CA125 (1:20; Dako), mesothelin (1:30; Novocastra), and pan cytokeratin (mixture of AE1/AE3 BM, 1:500, Dako; CAM5.2 Becton Dickinson, 1:50, San Jose, CA; cytokeratin MNF116 1:50, Carpinteria, CA; Keratin 8&18 1:25, Zymed, South San Francisco, CA).
Gene Expression Array and Data Analysis.
RNA was extracted with TRIZOL reagent (Invitrogen) and purified with the RNeasy Kit (Qiagen). The Affymetrix GeneChip Human Genome U133 series of oligoarrays (Affymetrix, Santa Clara, CA) were used to measure gene expression. This series tests the expression of >29,297 human genes and expressed sequence tags. Preparation of biotinylated cRNA, hybridization, and scanning of the microarrays were performed according to the manufacturer’s protocols. Data were collected by using GeneChip software (Affymetrix) and analyzed with the software program dChip (39
, 40)
. We used version 1.2 with the perfect match only model to estimate differences. The genes listed in Table 1
exhibited a 2.5-fold change and had to show consistent change in at least three of the six RAS-transformed (either HRASV12 or KRASV12) cell lines.
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B activity was measured by reporter plasmids that contained two copies of the wild-type NF-B binding site or two copies of mutated NF-B binding sites ligated to luciferase reporters. Transcriptional activation of interleukin (IL)-8 by NF-B was investigated by using a wild-type IL-8 promoter (-146 to +1) and a promoter that contained a mutated NF-B binding site (41)
. Luciferase activity was measured 48 h after transfection. A total of 4 µg of plasmid was used in each transfection. Activity was normalized against ß-galactosidase activity. Luciferase activity was assayed with Promega’s (Madison, WI) luciferase reporter assay system.
Preparation of Nuclear Extracts and Labeling of the Oligonucleotide and Electrophoretic Mobility Shift Assay.
Nuclear extract preparation was performed as described previously. Labeling of the oligonucleotide and electrophoretic mobility shift assay was performed according to manufacture’s instruction (Gelshift NF-B p50 kit from Active Motif Co., Carlsbad, CA; Refs. 41, 42, 43
).
Induction of Apoptosis by Antibody Treatment.
T29, T80, T29H, T29K, T80H, and T80K cells (2 x 105) were grown in serum-free RPMI medium for 24 h, after which, 2 µg/ml monoclonal antibody against IL-1ß or IL-8 (Sigma-Aldrich, MO) were added and the cells incubated without serum or antibiotics. Cells were harvested at 8, 16, 24, 48, and 72 h after the addition of antibody and stained with the Annexin V-fluorescence apoptosis detection kit I (Roche) and with propidium iodide, after which, cell apoptosis was analyzed with a FACStation (BD Biosciences) equipped with CellQuest software. The percentage of apoptotic cells was calculated in terms of peaks (M2) in the histogram, representing an early apoptotic population among the total cells analyzed.
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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. Telomerase activity was detected in T80 and T29 cells (Fig. 1, B and D
, Lane 3) but not in IOSE-80 or IOSE-29 cells (Fig. 1, B and D
, Lane 1). Heat inactivation eliminated the telomerase activity in both the T80 and T29 cell lines (Fig. 1, B and D
, Lane 4).
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and anchorage-independent cell growth (Table 2)
. The tumorigenicity of these ras-transformed human ovarian surface epithelial cells was evaluated by s.c. or i.p. injection into nude mice. No tumors were observed in the 2 mice injected with parental cells (IOSE-80 or IOSE-29) or in the 12 mice injected with immortalized cells T80 or 6 mice injected T29 cells (Table 2)
. By contrast, 5 of 6 mice given s.c. injections of T80H or T29H cells formed tumors with surface ulceration (Fig. 3A
, arrow), and 3 of 4 mice given i.p. injections of T80H cells developed ascites (Fig. 3, B and D)
or tumors that grew within the omentum (Fig. 3, C
, bottom arrow) or invaded the liver (Fig. 3, C
, top arrow). The average latency for s.c. tumor development was 21 days for T80H cells and 12 days for T29H cells.
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, numerous mitoses (Fig. 4B
, arrows), and extensive necrosis (Fig. 4C
, arrows). Tumor cells stained positively for cytokeratin (Fig. 4D)
, confirming their epithelial origin, and for WT-1 (Fig. 4E)
, a marker of serous ovarian carcinoma. The cells also stained positively for p53 (Fig. 4F)
, consistent with the presence of stabilized p53 protein induced by the SV40 T antigen. The tumors grown from T29H-inoculated cells showed morphology similar to high-grade sarcoma (data not shown). This is likely because of its parental cells, IOSE-29, which has elongated fibroblast morphology and did not stain with cytokeratin (35)
. The undifferentiated carcinomas and sarcoma are negative for CA125 and mesothelin (data not shown).
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. Injection of T29K cells produced only four s.c. tumors in 10 mice and one i.p. tumor in 4 mice (Table 2)
. Subcutaneous-injected T29K cells formed cystic structures in 3 of 6 mice within a week (Fig. 5B)
. Intraperitoneal-injected T29K cells produced a polyploid, cauliflower-shaped tumor (Fig. 5A)
that was undifferentiated (Fig. 5C)
and showed high nuclear pleomorphism (Fig. 5D
, arrow). Interestingly, peritoneal injection of T80K cells produced biphasic tumor with poorly differentiated carcinoma and sarcoma, similar to malignant mixed Mullerian tumor of ovary. The poorly differentiated carcinoma components invaded into adipose tissue (Fig. 5E)
and formed glandular-like structure (Fig. 5F
, arrows). Immunohistochemical staining of carcinoma cells are positive for cytokeratin (Fig. 5G)
, CA125 (Fig. 5H)
, and mesothelin (Fig. 5I)
.
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. The genes most consistently up-regulated were those for IL-1
, IL-1ß, IL-6, IL-8, IL-11, pre-B-cell colony-enhancing factor, prostaglandin-endoperoxide synthetase-2, Gro-1, and Gro-2. Other genes that were up-regulated mediate invasion and stromal remodeling (e.g., tissue factor inhibitor-2 and tissue inhibitors of matrix metalloproteinases 1 and 3). Down-regulated genes included those involved in stromal synthesis (e.g., collagen type III1 and type 12), the proapoptotic insulin-like growth factor binding protein-3, and the antiangiogenic factor thrombospondin-1.
RAS-Transformed Cells Depend on Elevated Cytokine for Survival.
To validate the changes in mRNA revealed by the expression array analysis, we used Western blotting to assess the amounts of protein produced by two of the up-regulated genes (those for IL-1ß and IL-8). IL-1ß and IL-8 were highly expressed in the RAS-transformed cells but not in the hTERT-immortalized T29 or T80 cells (Fig. 6)
. To examine the functional significance of this up-regulation in IL-1ß and IL-8 expression by RAS-mediated transformation, we treated immortalized cells and their RAS-transformed derivatives with monoclonal antibodies against IL-1ß and IL-8 and quantified the apoptotic cells. Numbers of apoptotic cells markedly increased when the RAS-transformed T29H and T80H cells were treated with these antibodies relative to similar treatment of the immortalized, nontransformed T29 and T80 cells (Fig. 7)
. Blocking IL-1ß in this way markedly increased the percentage of apoptotic cells in the T29H and T80H lines (Fig. 7, B and E)
; blocking IL-8 showed a similar effect (Fig. 7, B and E)
. The maximum effect of apoptosis is seen at 16–24 h, with longer incubations, and percentage of apoptotic cells is decreased, probably because of an optimal antibody and cytokine ratio at these time points. Antibodies against IL-1ß and IL-8 also induced apoptosis in KRAS-transformed cells, although not to the same extent as in the HRAS transformed cells (Fig. 7, C and F)
. These results may partially explain why the KRAS-transformed cells were less tumorigenic that the HRAS-transformed cells.
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, Lane 3), whereas no change was detected in SKOV-3 cells infected with either an empty retroviral vector (Fig. 8
, Lane 1) or a small interfering RNA against an HRAS with a mutated form of HRAS (Val12; Fig. 8
, Lane 2). These findings indicate that increased level of HRAS without V12 mutation plays a critical role in regulating cytokine expression, demonstrating that similar cytokine pathways are operative in a naturally derived ovarian cancer cell line as in our model.
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B.B in their promoter regions, we reasoned that NF-B could be mediating the cytokine expression in the RAS-transformed cells. To test this hypothesis, we used plasmids that contained two copies of a wild-type or a mutant NF-B binding site linked to a luciferase reporter. Incubating T29 and T80 cells with a retroviral expression vector for HRASV12 produced a concentration-dependent increase in luciferase activity from cells with the NF-B reporter constructs but not from those with the mutant NF-B binding-site controls (Fig. 9, A)
. Experiments with a plasmid containing the IL-8 promoter or an IL-8 promoter containing mutated NF-B binding sites produced similar results (Fig. 9B)
. However, when plasmids were transfected into T29H and T80H cells (i.e., those that already contain a stably expressed HRAS retroviral vector), high levels of luciferase activity were observed with wild-type NF-B binding sites but not with mutant NF-B binding sites. Cells without the stably integrated HRASV12 vector (T29 and T80) showed no elevation in luciferase activity (Fig. 9C)
. These results demonstrate that increased NF-B activity was associated with activation of IL-8.
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B in transforming activity of RAS, we used EMSA to directly examine the binding activity of NF-B. T29 cells showed basal levels of NF-B binding activity (Fig. 9D
, Lane 1) [the band at NF-B (p50)], similar to the levels produced by the positive control (Fig. 9D
, Lane 4); this NF-B binding activity could be competed away by oligonucleotide containing a wild-type NF-B binding site (Fig. 9D
, Lane 2) but not mutant oligonucleotide (Fig. 9D
, Lane 3). There was a marked increase in NF-B binding activity in the HRAS-transformed T29H cells (Fig. 9D
, Lane 5) that could be competed away by the oligonucleotide containing a wild-type NF-B binding site (Fig. 9D
, Lane 6) but not by an oligonucleotide containing the mutated site (Fig. 9D
, Lane 7). Similar results were found in T80 and T80H cells (Fig. 9D
, Lanes 9–16). These findings demonstrate that RAS transformation may be mediated through the transcription factor NF-B. |
DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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RAS exerts its effects through multiple downstream signal transduction pathways (45 , 46) . Analysis of changes in the expression of RAS-mediated genes can provide new insight into the pathways involved in RAS-mediated transformation. Previous studies have used NIH-3T3, an immortalized mouse fibroblast line or naturally occurring cancer cells to examine the mRNA changes associated with RAS-mediated transformation (47 , 48) . Extrapolating results from mice to humans, however, requires great caution; oncogenes that can transform rodent cells have repeatedly failed to transform human cells (49) , and RAS seems to use different mediators in transforming human cells and rodent cells (50) . In our study, a large fraction of the genes overexpressed as a consequence of HRAS or KRAS activation were for cytokines that are also involved in chronic inflammation and wound healing. Similarities between wound healing and tumor formation have long been hypothesized (51) . Increased concentrations of IL-1ß, IL-6, and IL-8 have been detected in human ovarian cancer cell lines (52) and in ascites and serum samples from patients with ovarian cancer (53, 54, 55, 56) , demonstrating that the pathways used in our genetically transformed cell lines are similar to those in naturally derived ovarian cancer. Furthermore, several of the up-regulated genes identified in our genetically transformed cells, including those superoxide dismutase 2, IL-6, tissue factor pathway inhibitor 2, and stanniocalcin 1, have also been identified independently by others (57) using expression array analysis of naturally derived human ovarian cancer tissue, demonstrating that our genetically defined model involves pathways that are dysregulated in naturally derived ovarian cancer.
Clinical features typical of ovarian cancer progression, including the spread of metastatic lesions over the peritoneal surface and the formation of "omental cake" that obstructs the bowel and invades the myenteric plexus, were mimicked in our model, as was the formation of ascites. Ascites can arise passively through occlusion of the diaphragmatic lymphatics by tumor cells, but nonobstructive mechanisms such as activation of a cytokine network in ras-mediated transformation may contribute as well. Our results demonstrate that NF-B was activated in ras-transformed ovarian epithelial cells and that this activation was responsible for the activation of IL-8, a regulator of angiogenesis, invasion, and metastasis and present in naturally derived cancer cell lines (58)
. NF-B has also been implicated in tumorigenesis (59
, 60)
, in suppression of the p53-independent apoptosis induced by oncogenic ras (61)
, and in HRAS-mediated transformation in rodent cells (41
, 61
, 63)
. Blockade of the NF-B pathway has been shown to decrease tumor growth in ovarian cancer and other cancer cell lines (40
, 42
, 64)
. These studies and our own work have demonstrated that both genetically transformed cells and naturally derived ovarian cancer cell lines use NF-B to activate cytokines, which facilitates the formation of tumor and ascites. Because ascites are associated with significant clinical morbidity, this model system may also be useful for testing strategies to limit malignant ascites.
In summary, we successfully transformed human ovarian surface epithelial cells with genetically defined elements that disrupt pathways that are often dysregulated in human ovarian cancer. Use of this system should permit identification of other oncogenic signals needed to transform normal ovarian epithelial cells, particularly these mimicked by the introduction of equivalent to SV40 T/t antigen, and allow the cooperative interaction of ovarian cancer genes to be tested in a genetically defined system. Understanding the molecular mechanisms of ras-mediated cytokine induction and their downstream targets will help to clarify the pathways in ovarian cancer development and help to identify new therapeutic and diagnostic targets.
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ACKNOWLEDGMENTS |
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B binding site and luciferase reporter constructs, and Dr. Suyun Huang for IL-8 promoter constructs. We also thank Janet E. Quinones for her expert help in immunohistochemical analysis. We thank the members of Jinsong Liu’s laboratory for helpful discussions, statistical analyses, and especially for Dr. Daniel Rosen for photographic assistance; Christine Wogan for excellent editorial assistance; and Dr. Zhibo Yang for his help in generating cell lines and initial tumorigenic assay. |
FOOTNOTES |
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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.
Requests for reprints: Dr. Jinsong Liu, Department of Pathology, Unit 85, The University of Texas. M. D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030. Phone: (713) 745-1102; Fax: (713) 792-5529; E-mail: jliu{at}mdanderson.org
Received 10/28/03. Revised 12/22/03. Accepted 12/23/03.
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REFERENCES |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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B signaling inhibits angiogenesis and tumorigenicity of human ovarian cancer cells by suppressing expression of vascular endothelial growth factor and interleukin 8. Cancer Res., 60: 5334-5339, 2000.B is required for H-ras oncogene induced abnormal cell proliferation and tumorigenesis. Oncogene, 19: 841-849, 2000.[CrossRef][Medline]
B activity by IBM suppresses tumorigenesis. Oncogene, 22: 1365-1370, 2003.[CrossRef][Medline]
in patients with epithelial ovarian cancer and their relationship to treatment with paclitaxel. Int. J. Gynecol. Cancer, 10: 33-41, 2000.
and ß genes. Cancer Res., 52: 2248-2252, 1992.B: control of oncogenesis and cancer therapy resistance. Biochim. Biophys. Acta, 1470: M55-M62, 2000.[Medline]
B in cancer: from innocent bystander to major culprit. Nat. Rev. Cancer, 2: 301-310, 2002.[CrossRef][Medline]
B activation to suppress p53-independent apoptosis induced by oncogenic Ras. Science (Wash. DC), 278: 1812-1815, 1997.B transcriptional activity, which is required for cellular transformation. J. Biol. Chem., 272: 24113-24116, 1997.B protects NIH 3T3 cells against stimulus-dependent apoptosis. Oncogene, 22: 477-483, 2003.[CrossRef][Medline]
B activity in human prostate cancer cells is associated with suppression of angiogenesis, invasion, and metastasis. Oncogene, 20: 4188-4197, 2001.[CrossRef][Medline]This article has been cited by other articles:
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Y. Hu, D. G. Rosen, Y. Zhou, L. Feng, G. Yang, J. Liu, and P. Huang Mitochondrial Manganese-Superoxide Dismutase Expression in Ovarian Cancer: ROLE IN CELL PROLIFERATION AND RESPONSE TO OXIDATIVE STRESS J. Biol. Chem., November 25, 2005; 280(47): 39485 - 39492. [Abstract] [Full Text] [PDF]
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A. Okamoto, T. Nikaido, K. Ochiai, S. Takakura, M. Saito, Y. Aoki, N. Ishii, N. Yanaihara, K. Yamada, O. Takikawa, et al. Indoleamine 2,3-Dioxygenase Serves as a Marker of Poor Prognosis in Gene Expression Profiles of Serous Ovarian Cancer Cells Clin. Cancer Res., August 15, 2005; 11(16): 6030 - 6039. [Abstract] [Full Text] [PDF]
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T. Daikoku, D. Wang, S. Tranguch, J. D. Morrow, S. Orsulic, R. N. DuBois, and S. K. Dey Cyclooxygenase-1 Is a Potential Target for Prevention and Treatment of Ovarian Epithelial Cancer Cancer Res., May 1, 2005; 65(9): 3735 - 3744. [Abstract] [Full Text] [PDF]
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K. W. Cheng, J. P. Lahad, J. W. Gray, and G. B. Mills Emerging Role of RAB GTPases in Cancer and Human Disease Cancer Res., April 1, 2005; 65(7): 2516 - 2519. [Abstract] [Full Text] [PDF]
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 M. A. Spillman and A. Berchuck Stretching the Surface Epithelium Reproductive Sciences, December 1, 2004; 11(8): 501 - 502. [PDF]
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T. W. Young, F. C. Mei, G. Yang, J. A. Thompson-Lanza, J. Liu, and X. Cheng Activation of Antioxidant Pathways in Ras-Mediated Oncogenic Transformation of Human Surface Ovarian Epithelial Cells Revealed by Functional Proteomics and Mass Spectrometry Cancer Res., July 1, 2004; 64(13): 4577 - 4584. [Abstract] [Full Text] [PDF]
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