- Research article
- Open Access
Effect of PTTG on endogenous gene expression in HEK 293 cells
© Panguluri and Kakar; licensee BioMed Central Ltd. 2009
- Received: 6 July 2009
- Accepted: 3 December 2009
- Published: 3 December 2009
Pituitary tumor transforming gene (PTTG), also known as securin, is highly expressed in various tumors including pituitary, thyroid, colon, ovary, testis, lung, and breast. An overexpression of PTTG enhances cell proliferation, induces cellular transformation in vitro, and promotes tumor development in nude mice. PTTG also inhibits separation of sister chromatids leading to aneuploidy and genetic instability. A great amount of work has been undertaken to understand the biology of PTTG and its expression in various tumors. However, mechanisms by which PTTG mediates its tumorigenic function are not fully understood. To utilize this gene for cancer therapy, identification of the downstream signaling genes regulated by PTTG in mediation of its tumorigenic function is necessary. For this purpose, we expressed PTTG in human embryonic kidney (HEK293) cells that do not express PTTG and analyzed the downstream genes using microarray analysis.
A total of 22,277 genes printed on an Affymetrix HG-U133A 2.0 GeneChip™ array were screened with labeled cRNA prepared from HEK293 cells infected with adenovirus vector expressing PTTG cDNA (AdPTTG cDNA) and compared with labeled cRNA prepared from HEK293 cells infected with control adenovirus (control Ad) or adenovirus vector expressing GFP (AdGFP). Out of 22,277 genes, 71 genes were down-regulated and 35 genes were up-regulated with an FDR corrected p-value of ≤ 0.05 and a fold change of ≥2. Most of the altered genes identified are involved in the cell cycle and cell apoptosis; a few are involved in mRNA processing and nitrogen metabolism. Most of the up-regulated genes belong to the histone protein family.
PTTG is a well-studied oncogene for its role in tumorigenesis. In addition to its importance in regulation of the cell cycle, this gene has also been recently shown to play a role in the induction of cell apoptosis. The microarray analysis in the present study demonstrated that PTTG may induce apoptosis by down-regulation of oncogenes such as v-Jun and v-maf and up-regulation of the histone family of genes.
Pituitary tumor transforming gene (PTTG) is a multifunctional protein involved in the cell cycle, cell proliferation, angiogenesis, metastasis, and other cellular functions. PTTG was initially isolated from rat pituitary tumor . Based on its function in the inhibition of separation of sister chromatids, it was named securin . Overexpression of PTTG was reported in many cancers including ovary, lung, testis, kidney, colon, thyroid, pituitary, liver, adrenal, breast, prostate, melanoma, leukemia, and lymphoma [3–15]. Many laboratories have explored this oncogene for its role in tumorigenesis since its identification (see reference  for review). Pei and Melmed  showed that overexpression of PTTG in NIH3T3 cells inhibits cell proliferation and induces cell transformation in vitro. In contrast, overexpression of PTTG in human embryonic kidney (HEK293) cells was shown to increase cell proliferation, induce cellular transformation in vitro, and promote tumor development in nude mice [7, 17]. Ishikawa et al.  and McCabe et al.  showed PTTG's role in angiogenesis by coinciding its function in inducing the expression of basic fibroblast growth factor (bFGF) and vascular endothelial cell growth factor (VEGF). Heaney et al.  reported regulation of PTTG expression in vitro and in vivo by estrogen. Ishikawa et al.  demonstrated that the conditioned medium collected from NIH3T3 cells overexpressing human PTTG induced angiogenesis through up-regulation of bFGF both in vitro and in vivo. This information was further confirmed by Kim et al.  who showed regulation of angiogenic genes such as ID3 and TSP-1 by PTTG. Angiogenic gene ID3 has been shown to be up-regulated by VEGF , which is believed to play a critical role in cell proliferation and to be a precursor of endothelial cell recruitment [23, 24]. Shibata et al.  showed the importance of PTTG in tumor metastasis and its correlation with the pathological stage, levels of pain, and extensive lymph node metastasis in esophageal cancer patients. Its expression levels were found to be correlated with a higher degree of tumor recurrence and tumor aggression in breast cancer as well as in squamous cell carcinoma [26, 27]. In our previous studies, we  demonstrated the up-regulation of matrix metalloproteinase (MMP)-2 and its relationship in tumor angiogenesis and metastasis.
Role of PTTG in tumorigenesis was further confirmed by down-regulation of PTTG in tumors. Chen et al down-regulated PTTG expression using full-length anti-PTTG complementary (c) DNA in SKOV3 and showed a significant reduction of cell proliferation and colony formation on soft agar compared to the non-transformed cells and bFGF protein levels. In this context, using PTTG siRNA to transfect an ovarian tumor cell line (A2780), El-Naggar et al.  showed a 50% reduction in cell proliferation and a 70% reduction in colony formation. In addition, these investigators demonstrated a reduced incidence of tumor development and tumor growth in nude mice injected with A2780 cells that constitutively expressed PTTG siRNA compared to cells expressing control siRNA. These results were confirmed by Cho-Rok et al.  using an adenovirus expression system expressing PTTG siRNA in a hapotoma cell line.
The molecular mechanisms by which PTTG achieves its functions remain unclear. Although a limited number of individual target genes in various cell lines have been identified, a large-scale profile of the target genes regulated by PTTG had not been undertaken at the time of this study. For this purpose, we explored the cDNA-microarray technology to identify the differentially expressed genes by PTTG. Most of the tumors and cancer cell lines overexpress PTTG; therefore, we selected HEK293 cells, which expresses a very low or undetectable level of endogenous PTTG. The change in expression of some genes by PTTG identified by cDNA microarray analysis was confirmed by quantitative real-time PCR (qRT-PCR).
HEK293 cells were propagated in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) according to instructions from American Type Culture Collection (ATCC; Manassas, VA). Medium and serum were purchased from Invitrogen (Carlsbad, CA). Cells were grown under 5% CO2 at 37°C.
The full length PTTG cDNA was sub-cloned into adenovirus shuttle vector pShuttle. Positive clones were sequenced to confirm the sequence and orientation of cDNA. The adenovirus expression system was generated and purified in association with the Gene Therapy Center, Virus Vector Core Facility, University of North Carolina at Chapel Hill.
Expression of PTTG in HEK293 cells using the adenovirus expression system
HEK293 cells growing in log phase were trypsinized and plated on T-75 flasks. Cells were infected with adenovirus vector, adenovirus vector expressing GFP (AdGFP), or adenovirus vector expressing PTTG cDNA (AdPTTG cDNA) at variable multiplicities of infection (MOI). Infection of cells was carried out in serum-free DMEM medium for 2 h. After 2 h, the medium was replaced with regular growth medium and incubated at 37°C under 5% CO2. The optimum MOI was found to be 1:10 to provide a 90- to 95% level of infection. Experiments were performed in accordance with instructions from the University of Louisville Institutional Biosafety Committee.
Isolation of total RNA
After 48 h of post-infection with adenovirus at a MOI of 1:10, medium was aspirated and cells were rinsed twice with phosphate-buffered saline (PBS). Cells were harvested by scraping with a rubber policeman and centrifuged at 5000 rpm for 5 min. The medium was discarded and 1 ml of Triazol reagent (Sigma-Aldrich, St. Louis, MO) was added to the cell pellet. The cells were homogenized and total RNA was purified as described previously . Quality of RNA was examined on agarose gel electrophoresis and quantified by NanoDrop 1000 (Thermo Scientific, Waltham, MA). All treatments were performed in triplicate.
Western blot analysis
Cells were cultured in complete growth media to 80-90% confluence. Cells were washed and collected in cold PBS and lysed with lysis buffer (10 mM Tris-HCl, pH 7.5; 150 mM NaCl; 1% Triton X-100; 1 mM EDTA). Protein concentration for each sample was assayed by Bradford Method (Bio-Rad Laboratories). Forty micrograms of protein from each sample was resolved on 15% SDS polyacrylamide gel and transferred to nitrocellulose membranes (GE Healthcare Biosciences, Piscataway, NJ). Membranes were blocked overnight with 5% non-fat milk/Tween (20 mM Tris/HCl, 150 mM NaCl, 0.1% Tween-20, pH 7.6) (TBST). Membranes were incubated with PTTG antiserum diluted at (1:1,500)  for 1 h at room temperature followed by 3 washes with TBST and then incubated with HRP-conjugated goat anti-rabbit secondary antibody diluted at 1:5,000. The immune complexes formed were detected by ECL reagents (GE Healthcare Biosciences). The blots were stripped and reprobed with β-actin antibody (Sigma-Aldrich, USA.) to examine equal loading.
The microarray analysis was performed at the University of Louisville Microarray Core Facility according to instructions from Affymetrix (Santa Clara, CA). mRNA was converted into double stranded cDNA using a T7-oligo (dT) promoter primer sequence. The double-stranded cDNA was purified and served as a template in the subsequent in vitro transcription reactions. The in vitro transcription reactions were carried out in the presence of T7 RNA polymerase and a biotinylated nucleotide analog/ribonucleotide mix for cRNA amplification. The biotinylated cRNA was purified, fragmented, and used in the hybridization cocktail containing control oligonucleotide B2 and four control bacterial and phage cDNA (BioB, BioC, BioD, cre). The labeled cRNA was hybridized to GeneChip expression arrays Human U133A 2.0 (Affymetrix), containing 22,277 genes using the protocol described by Affymetrix and as described previously . Alterations in RNA transcript levels (AdPTTG cDNA vs. adenovirus or adenovirus vs. AdGFP) were analyzed using Partek Genomics Suite 6.2 (Partek Inc., St. Louis, MO). Three different experiments were performed for statistical analysis.
Data analysis was performed using Partek Genomics Suite 6.2 (Partek Inc., St. Louis, MO). The Affymetrix probe level signal values were summarized using the RMA algorithm. Statistically significant changed genes were identified by analysis of variance (ANOVA) with FDR-corrected p-values < 0.05. The contrast between adenovirus vs. AdGFP yielded no significantly different in genes based on these parameters. Two-way ANOVA tests were carried out to identify differentially expressed genes in the comparison of AdPTTG cDNA vs. adenovirus, taking treatment and batch effect for the triplicate sample processing into account. The genes that showed 2-fold induction or 2-fold suppression were transferred to separate up and down lists, respectively. The gene sets with an FDR corrected p-value of less than 0.05 were identified in these lists and Ingenuity Pathway Analysis software (Ingenuity Systems Inc., Redwood City, CA) was used to interpret the interactive pathway networks between the selected genes from the microarray data.
The genes that showed signal detection values more than 100, p-values less than 0.001, and a change in gene expression of more than 2-fold when compared to the control were selected to study their relative expressions by qRT-PCR using a MyiQ single color real-time PCR detection system (Bio-Rad Laboratories, Hercules, CA). qRT-PCR was performed in a total of 20 μl reaction volume containing 1 μl of cDNA, 1 μl each of forward- and reverse-specific primers (from 10 μM primer stock), 10 μl of supermix (Bio-Rad Laboratories), and 7 μl of nuclease-free water. All qRT-PCR reactions were performed using the following reaction conditions: initial denaturation at 95°C for 3 min followed by 45 cycles at 95°C for 10 s, 60°C for 20 s, and 72°C for 30 s. A fluorescence reading determined the extent of amplification at the end of each cycle.
The primer sequences used for the validation of differentially expressed genes by PTTG overexpression through qRT-PCR.
5' gccttagatgggagatctca 3'
5' gctttaacagtcttctcagt 3'
5' gcgtgcgctcttagagaaac 3'
5' ccgttgctggactggattat 3'
5' cccgaccgaacagaagac 3'
5' gcttggtgatgatggtgatg 3'
5' ccgcaaagagagctactcca 3'
5' ggagctggtgtacttggtga 3'
5' agcgtagcggagtttctctg 3'
5' tagcgctcttcttcggagtt 3'
5' gcagccgcaaagagacttac 3'
5' tggagctggtgtacttggtg 3'
5' agaagaacaacagccgcatc 3'
5' gcttcttcgccttctttgg 3'
Results and Discussion
Adenovirus infection showed 90% efficiency
PTTG overexpression caused down-regulation of many genes
The differentially expressed genes with different p-values in PTTG-expressing HEK293 cells by microarray analysis.
v-Jun and Maf were down-regulated in PTTG-expressed HEK293 cells
The present study also showed the down-regulation of another leucine zipper-bearing transcriptional factor, v-Maf, by 4-fold in microarray data, which is consistent with qRT-PCR analysis (Fig. 3 and 4). The recent findings also suggest that mutated Jun, which lacks a transactivation domain, represses cell transformation not only by itself, but also by another oncogene, Maf . From their studies, these investigators suggest that these two oncogenes share downstream targets and induce cell transformation through similar genes. Our studies showed a correlation with these findings as both of these genes down-regulated in microarray as well as in qRT-PCR. The ingenuity pathway analysis showed that members of Maf gene transcriptional factors regulate insulin gene transcription in islet cells . In turn, this was supported by the findings of Wang et al. , who showed that PTTG knockout mice severely impair glucose homeostasis leading to diabetes during late adulthood due to impaired beta cell proliferation. From these observations, it is evident that PTTG has its role in the cell cycle and tumorigenesis, but also is involved in maintaining glucose homeostasis.
Histone family of genes was up-regulated by PTTG overexpression
PTTG is a well-known oncogene, which is found to be expressed in many tumors and tumor cell lines. Existing evidence shows that this gene is involved in cell proliferation, migration, tumor invasion, and metastasis. In addition to these tumorigenic functions, this gene plays an important role in cell division. The recent findings of PTTG in inducing apoptosis  brings forth a new role for this gene in causing apoptosis rather than inducing cell proliferation and tumorigenesis. To identify the differentially expressed genes by PTTG overexpression in HEK293 cells, we have explored the oligonucleotide-microarray and qRT-PCR techniques. Our data clearly indicate the down-regulation of v-Maf and Jun and the up-regulation of many histone genes, which may explain the pathways by which PTTG induces apoptotic genes. In addition, the fate of the cell that overexpresses PTTG to choose either a tumorigenic or an apoptotic pathway may be based on its quantity of expression, which reveals this gene as playing dual roles.
Part of this work was performed with assistance of the University of Louisville Microarray Facility, which is supported by NCRR COBRE P20RR018733, KY-INBRE NCRR P20RR016481 and the James Graham Brown Cancer Center at University of Louisville. Authors are thankful to Mr. Andrew Mash for critical editing of the manuscript. This work was supported by a grant from NCI CA124630 (SSK).
- Pei L, Melmed S: Isolation and characterization of a pituitary tumor-transforming gene (PTTG). Molecular endocrinology (Baltimore, Md). 1997, 11 (4): 433-441. 10.1210/me.11.4.433.View ArticleGoogle Scholar
- Zou H, McGarry TJ, Bernal T, Kirschner MW: Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis. Science. 1999, 285 (5426): 418-422. 10.1126/science.285.5426.418.View ArticlePubMedGoogle Scholar
- Ogbagabriel S, Fernando M, Waldman F, Bose S, Heaney A: Securin is overexpressed in breast cancer. Mod Pathol. 2005, 18: 985-990. 10.1038/modpathol.3800382.View ArticlePubMedGoogle Scholar
- Yu R, Ren S, Horwitz G, Wang Z, Melmed S: Pituitary tumor transforming gene (PTTG) regulates placental JEG-3 cell division and survival: evidence from live cell imaging. Mol Endocrinol. 2000, 14: 1137-1146. 10.1210/me.14.8.1137.View ArticlePubMedGoogle Scholar
- Puri R, Tousson A, Chen L, Kakar S: Molecular cloning of pituitary tumor transforming gene 1 from ovarian tumors and its expression in tumors. Cancer Lett. 2001, 163: 131-139. 10.1016/S0304-3835(00)00688-1.View ArticlePubMedGoogle Scholar
- Dominguez A, Ramos-Morales F, Romero F, Rios R, Dreyfus F, Tortolero M, Pintor-Toro J: hpttg, a human homologue of rat pttg, is overexpressed in hematopoietic neoplasms. Evidence for a transcriptional activation function of hPTTG. Oncogene. 1998, 17: 2187-2193. 10.1038/sj.onc.1202140.View ArticlePubMedGoogle Scholar
- Kakar S, Jennes L: Molecular cloning and characterization of the tumor transforming gene (TUTR1): a novel gene in human tumorigenesis. Cytogenet Cell Genet. 1999, 84: 211-216. 10.1159/000015261.View ArticlePubMedGoogle Scholar
- Heaney A, Nelson V, Fernando M, Horwitz G: Transforming events in thyroid tumorigenesis and their association with follicular lesions. J Clin Endocrinol Metab. 2001, 86: 5025-5032. 10.1210/jc.86.10.5025.View ArticlePubMedGoogle Scholar
- Ramos-Morales F, Dominguez A, Romero F, Luna R, Multon M, Pintor-Toro J, Tortolero M: Cell cycle regulated expression and phosphorylation of hpttg proto-oncogene product. Oncogene. 2000, 19: 403-409. 10.1038/sj.onc.1203320.View ArticlePubMedGoogle Scholar
- Zhang X, Horwitz GA, Prezant TR, Valentini A, Nakashima M, Bronstein MD, Melmed S: Structure, expression, and function of human pituitary tumor-transforming gene (PTTG). Mol Endocrinol. 1999, 13 (1): 156-166. 10.1210/me.13.1.156.View ArticlePubMedGoogle Scholar
- Heaney A, Singson R, McCabe C, Nelson V, Nakashima M, Melmed S: Expression of pituitary-tumour transforming gene in colorectal tumours. Lancet. 2000, 26: 716-719. 10.1016/S0140-6736(99)10238-1.View ArticleGoogle Scholar
- Ramaswamy S, Ross K, Lander E, Golub T: A molecular signature of metastasis in primary solid tumors. Nature Genetics. 2003, 33: 49-54. 10.1038/ng1060.View ArticlePubMedGoogle Scholar
- Zhang X, Horwitz G, Heany A, Nakashima M, Prezant T, Bronstein M, Melmed S: Pituitary tumor transforming gene (PTTG) expression in pituitary adenomas. J Clin Endocrinol Metab. 1999, 84: 761-767. 10.1210/jc.84.2.761.View ArticlePubMedGoogle Scholar
- Heaney A, Fernando M, Melmed S: Functional role of estrogen in pituitary tumor pathogenesis. J Clin Invest. 2002, 109: 277-283.PubMed CentralView ArticlePubMedGoogle Scholar
- Boelaert K, McCabe C, Tannahill L, Gittoes N, Holder R, Watkinson J, Bradwell A, Sheppard M, Franklyn J: Pituitary tumor transforming gene and fibroblast growth factor-2 expression: potential prognostic indicators in differential thyroid cancer. J Clin Endocr Metab. 2003, 88: 2341-2347. 10.1210/jc.2002-021113.View ArticlePubMedGoogle Scholar
- Panguluri S, Yeakel C, Kakar S: PTTG: an important target gene for ovarian cancer therapy. Journal of Ovarian Research. 2008, 1 (1): 6-10.1186/1757-2215-1-6.PubMed CentralView ArticlePubMedGoogle Scholar
- Hamid T, Kakar S: PTTG/securin activates expression of p53 and modulates its function. Mol Cancer. 2004, 3: 18-10.1186/1476-4598-3-18.PubMed CentralView ArticlePubMedGoogle Scholar
- Ishikawa H, Heaney A, Yu R, Horwitz G, Melmed S: Human pituitary tumor-transforming gene induces angiogenesis. J Clin Endocrinol Metab. 2001, 86: 867-874. 10.1210/jc.86.2.867.PubMedGoogle Scholar
- McCabe C, Boelaert K, Tannahill L, Heaney A, Stratford A, Khaira J, Hussain S, Sheppard M, Franklyn J, Gittoes N: Vascular endothelial growth factor, its receptor KDR/Flk-1, and pituitary tumor transforming gene in pituitary tumors. J Clin Endocrinol Metab. 2002, 87: 4238-4244. 10.1210/jc.2002-020309.View ArticlePubMedGoogle Scholar
- Heaney A, Horwitz G, Wang Z, Singson R, Melmed S: Early involvement of estrogen-induced pituitary tumor transforming gene and fibroblast growth factor expression in prolactinoma pathogenesis. Nat Med. 1999, 5: 1317-1321. 10.1038/15275.View ArticlePubMedGoogle Scholar
- Kim D, Buchanan M, Stratford A, Watkinson J, Eggo M, Franklyn J, McCabe C: PTTG promotes a novel VEGF-KDR-ID3 autocrine mitogenic pathway in thyroid cancer. Clin Otolaryngol. 2006, 31: 246-10.1111/j.1749-4486.2006.01236_6.x.View ArticleGoogle Scholar
- Lyden D, Young AZ, Zagzag D, Yan W, Gerald W, O'Reilly R, Bader BL, Hynes RO, Zhuang Y, Manova K, et al: Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts. Nature. 1999, 401 (6754): 670-677. 10.1038/44334.View ArticlePubMedGoogle Scholar
- Lasorella A, Uo T, Iavarone A: Id proteins at the cross-road of development and cancer. Oncogene. 2001, 20: 8326-8333. 10.1038/sj.onc.1205093.View ArticlePubMedGoogle Scholar
- Lyden D, Hattori K, Dias S, Costa C, Blaikie P, Butros L, Chadburn A, Heissig B, Marks W, Witte L, et al: Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med. 2001, 7: 1194-1201. 10.1038/nm1101-1194.View ArticlePubMedGoogle Scholar
- Shibata Y, Haruki N, Kuwabara Y, Nishiwaki T, Kato J, Shinoda N, Sato A, Kimura M, Koyama H, Toyama T: Expression of PTTG (pituitary tumor transforming gene) in esophageal cancer. Jpn J Clin Oncol. 2002, 32: 233-237. 10.1093/jjco/hyf058.View ArticlePubMedGoogle Scholar
- Solbach C, Roller M, Fellbaum C, Nicoletti M, Kaufmann M: PTTG mRNA expression in primary breast cancer: a prognostic marker for lymph node invasion and tumor recurrence. Breast (Edinburgh, Scotland). 2004, 13 (1): 80-81.View ArticleGoogle Scholar
- Solbach C, Roller M, Eckerdt F, Peters S, Knecht R: Pituitary tumor-transforming gene expression is a prognostic marker for tumor recurrence in squamous cell carcinoma of the head and neck. BMC cancer. 2006, 6: 242-10.1186/1471-2407-6-242.PubMed CentralView ArticlePubMedGoogle Scholar
- Malik M, Kakar S: Regulation of angiogenesis and invasion by human Pituitary tumor transforming gene (PTTG) through increased expression and secretion of matrix metalloproteinase-2 (MMP-2). Mol Cancer. 2006, 5: 61-10.1186/1476-4598-5-61.PubMed CentralView ArticlePubMedGoogle Scholar
- Chen G, Li J, Li F, Li X, Zhou J, Lu Y, Ma D: Inhibitory effects of anti-sense PTTG on malignant phenotype of human ovarian carcinoma cell line SK-OV-3. J Huazhong Univ Sci Technolog Med Sci. 2004, 24 (4): 369-372. 10.1007/BF02861870.View ArticlePubMedGoogle Scholar
- El-Naggar S, Malik M, Kakar S: Small interfering RNA against PTTG: a novel therapy for ovarian cancer. Int J Oncol. 2007, 31: 137-143.PubMedGoogle Scholar
- Cho-Rok J, Yoo J, Jang Y, Kim S, Chu I, Yeom Y, Choi J, Im D: Adenovirus-mediated transfer of siRNA against PTTG1 inhibits liver cancer cell growth in vitro and in vivo. Hepatology. 2006, 43: 1042-1052. 10.1002/hep.21137.View ArticlePubMedGoogle Scholar
- Kakar S, Chen L, Puri R, Flynn S, Jennes L: Characterization of a polyclonal antibody to human pituitary tumor transforming gene 1 (PTTG1) protein. J Histochem Cytochem. 2001, 49: 1537-1546.View ArticlePubMedGoogle Scholar
- Panguluri SK, Li B, Hormann RE, Palli SR: Effect of ecdysone receptor gene switch ligands on endogenous gene expression in 293 cells. The FEBS journal. 2007, 274 (21): 5669-5689. 10.1111/j.1742-4658.2007.06089.x.View ArticlePubMedGoogle Scholar
- Maki Y, Bos TJ, Davis C, Starbuck M, Vogt PK: Avian sarcoma virus 17 carries the jun oncogene. Proceedings of the National Academy of Sciences of the United States of America. 1987, 84 (9): 2848-2852. 10.1073/pnas.84.9.2848.PubMed CentralView ArticlePubMedGoogle Scholar
- Vogt PK, Bos TJ: The oncogene jun and nuclear signalling. Trends in biochemical sciences. 1989, 14 (5): 172-175. 10.1016/0968-0004(89)90268-5.View ArticlePubMedGoogle Scholar
- Shaulian E, Schreiber M, Piu F, Beeche M, Wagner EF, Karin M: The mammalian UV response: c-Jun induction is required for exit from p53-imposed growth arrest. Cell. 2000, 103 (6): 897-907. 10.1016/S0092-8674(00)00193-8.View ArticlePubMedGoogle Scholar
- Maclaren A, Clark W, Black EJ, Gregory D, Fujii H, Gillespie DA: v-Jun stimulates both cdk2 kinase activity and G1/S progression via transcriptional repression of p21 CIP1. Oncogene. 2003, 22 (16): 2383-2395. 10.1038/sj.onc.1206329.View ArticlePubMedGoogle Scholar
- El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B: WAF1, a potential mediator of p53 tumor suppression. Cell. 1993, 75 (4): 817-825. 10.1016/0092-8674(93)90500-P.View ArticlePubMedGoogle Scholar
- Mu YM, Oba K, Yanase T, Ito T, Ashida K, Goto K, Morinaga H, Ikuyama S, Takayanagi R, Nawata H: Human pituitary tumor transforming gene (hPTTG) inhibits human lung cancer A549 cell growth through activation of p21(WAF1/CIP1). Endocrine journal. 2003, 50 (6): 771-781. 10.1507/endocrj.50.771.View ArticlePubMedGoogle Scholar
- Leaner VD, Kinoshita I, Birrer MJ: AP-1 complexes containing cJun and JunB cause cellular transformation of Rat1a fibroblasts and share transcriptional targets. Oncogene. 2003, 22 (36): 5619-5629. 10.1038/sj.onc.1206644.View ArticlePubMedGoogle Scholar
- Jogi A, Persson P, Grynfeld A, Pahlman S, Axelson H: Modulation of basic helix-loop-helix transcription complex formation by Id proteins during neuronal differentiation. The Journal of biological chemistry. 2002, 277 (11): 9118-9126. 10.1074/jbc.M107713200.View ArticlePubMedGoogle Scholar
- Tsai SJ, Lin SJ, Cheng YM, Chen HM, Wing LY: Expression and functional analysis of pituitary tumor transforming gene-1 [corrected] in uterine leiomyomas. The Journal of clinical endocrinology and metabolism. 2005, 90 (6): 3715-3723. 10.1210/jc.2004-2303.View ArticlePubMedGoogle Scholar
- Pollmann C, Huang X, Mall J, Bech-Otschir D, Naumann M, Dubiel W: The constitutive photomorphogenesis 9 signalosome directs vascular endothelial growth factor production in tumor cells. Cancer research. 2001, 61 (23): 8416-8421.PubMedGoogle Scholar
- Hamid T, Malik M, Kakar S: Ectopic expression of PTTG1/securin promotes tumorigenesis in human embryonic kidney cells. Mol Cancer. 2005, 4: 3-10.1186/1476-4598-4-3.PubMed CentralView ArticlePubMedGoogle Scholar
- Kim DS, Franklyn JA, Boelaert K, Eggo MC, Watkinson JC, McCabe CJ: Pituitary tumor transforming gene (PTTG) stimulates thyroid cell proliferation via a vascular endothelial growth factor/kinase insert domain receptor/inhibitor of DNA binding-3 autocrine pathway. J Clin Endocrinol Metab. 2006, 91 (11): 4603-4611. 10.1210/jc.2006-1291.View ArticlePubMedGoogle Scholar
- Heaney A, Fernando M, Melmed S: Functional role of estrogen in pituitary tumor pathogenesis. J Clin Invest. 2002, 109: 277-28399.PubMed CentralView ArticlePubMedGoogle Scholar
- Kataoka K, Shioda S, Yoshitomo-Nakagawa K, Handa H, Nishizawa M: Maf and Jun nuclear oncoproteins share downstream target genes for inducing cell transformation. The Journal of biological chemistry. 2001, 276 (39): 36849-36856. 10.1074/jbc.M102234200.View ArticlePubMedGoogle Scholar
- Matsuoka TA, Zhao L, Artner I, Jarrett HW, Friedman D, Means A, Stein R: Members of the large Maf transcription family regulate insulin gene transcription in islet beta cells. Molecular and cellular biology. 2003, 23 (17): 6049-6062. 10.1128/MCB.23.17.6049-6062.2003.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang Z, Moro E, Kovacs K, Yu R, Melmed S: Pituitary tumor transforming gene-null male mice exhibit impaired pancreatic beta cell proliferation and diabetes. Proceedings of the National Academy of Sciences of the United States of America. 2003, 100 (6): 3428-3432. 10.1073/pnas.0638052100.PubMed CentralView ArticlePubMedGoogle Scholar
- Geer van der P, Hunter T, Lindberg RA: Receptor protein-tyrosine kinases and their signal transduction pathways. Annual review of cell biology. 1994, 10: 251-337. 10.1146/annurev.cb.10.110194.001343.View ArticlePubMedGoogle Scholar
- Catalano A, Rodilossi S, Rippo MR, Caprari P, Procopio A: Induction of stem cell factor/c-Kit/slug signal transduction in multidrug-resistant malignant mesothelioma cells. The Journal of biological chemistry. 2004, 279 (45): 46706-46714. 10.1074/jbc.M406696200.View ArticlePubMedGoogle Scholar
- Murphy LO, MacKeigan JP, Blenis J: A network of immediate early gene products propagates subtle differences in mitogen-activated protein kinase signal amplitude and duration. Molecular and cellular biology. 2004, 24 (1): 144-153. 10.1128/MCB.24.1.144-153.2004.PubMed CentralView ArticlePubMedGoogle Scholar
- Maeyama M, Koga H, Selvendiran K, Yanagimoto C, Hanada S, Taniguchi E, Kawaguchi T, Harada M, Ueno T, Sata M: Switching in discoid domain receptor expressions in SLUG-induced epithelial-mesenchymal transition. Cancer. 2008, 113 (10): 2823-2831. 10.1002/cncr.23900.View ArticlePubMedGoogle Scholar
- Vitali R, Mancini C, Cesi V, Tanno B, Mancuso M, Bossi G, Zhang Y, Martinez RV, Calabretta B, Dominici C, et al: Slug (SNAI2) down-regulation by RNA interference facilitates apoptosis and inhibits invasive growth in neuroblastoma preclinical models. Clin Cancer Res. 2008, 14 (14): 4622-4630. 10.1158/1078-0432.CCR-07-5210.View ArticlePubMedGoogle Scholar
- Radic M, Marion T, Monestier M: Nucleosomes Are Exposed at the Cell Surface in Apoptosis. 2004, 172: 6692-6700.Google Scholar
- Cheung WL, Briggs SD, Allis CD: Acetylation and chromosomal functions. Curr Opin Cell Biol. 2000, 12: 326-333. 10.1016/S0955-0674(00)00096-X.View ArticlePubMedGoogle Scholar
- Mahadevan LC, Willis AC, Barratt MJ: Rapid histone H3 phosphorylation in response to growth factors, phorbol esters, okadaic acid, and protein synthesis inhibitors. Cell. 1991, 65: 775-783. 10.1016/0092-8674(91)90385-C.View ArticlePubMedGoogle Scholar
- Sassone-Corsi P, Mizzen CA, Cheung P, Crosio C, Monaco L, Jacquot S: Requirement of Rsk-2 for epidermal growth factor-activated phosphorylation of histone H3. Science. 1999, 285: 886-891. 10.1126/science.285.5429.886.View ArticlePubMedGoogle Scholar
- Chen SL, Loffler KA, Chen D, Stallcup MR, Muscat GE: The coactivator-associated arginine methyltransferase is necessary for muscle differentiation: CARM1 coactivates myocyte enhancer factor-2. J Biol Chem. 2002, 277: 4324-4333. 10.1074/jbc.M109835200.View ArticlePubMedGoogle Scholar
- Beaujean N: Fundamental features of chromatin structure. Cloning Stem Cells. 2002, 4: 355-361. 10.1089/153623002321025032.View ArticlePubMedGoogle Scholar
- Konishi A, Shimizu S, Hirota J, Takao T, Fan Y, Matsuoka Y, Zhang L, Yoneda Y, Fujii Y, Skoultchi AI, et al: Involvement of histone H1.2 in apoptosis induced by DNA double-strand breaks. Cell. 2003, 114 (6): 673-688. 10.1016/S0092-8674(03)00719-0.View ArticlePubMedGoogle Scholar
- Pagel P, Kovac S, Oesterheld M, Brauner B, Dunger-Kaltenbach I, Frishman G, Montrone C, Mark P, Stumpflen V, Mewes HW, et al: The MIPS mammalian protein-protein interaction database. Bioinformatics (Oxford, England). 2005, 21 (6): 832-834. 10.1093/bioinformatics/bti115.View ArticleGoogle Scholar
- Hall C, Nelson DM, Ye X, Baker K, DeCaprio JA, Seeholzer S, Lipinski M, Adams PD: HIRA, the human homologue of yeast Hir1p and Hir2p, is a novel cyclin-cdk2 substrate whose expression blocks S-phase progression. Mol Cell Biol. 2001, 21 (5): 1854-1865. 10.1128/MCB.21.5.1854-1865.2001.PubMed CentralView ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.