The online version of this article (doi:10.1186/1477-7819-10-183) contains supplementary material, which is available to authorized users.
Tiandong Han, Donghao Shang contributed equally to this work.
The authors declare that they have no competing interests.
TH, DS conceived and designed the experiments. TH, XX performed the experiments. DS analyzed the data. DS, XX contributed reagents/materials/ analysis tools. TH, YT drafted the manuscript. All authors read and approved the final manuscript.
Renal cell carcinoma (RCC) is one of the most common kidney cancers and is highly resistant to chemotherapy. We previously demonstrated that 5-aza-2′-deoxycytidine (DAC) could significantly increase the susceptibility of renal cell carcinoma (RCC) cells to paclitaxel (PTX) treatment in vitro, and showed the synergy of DAC and PTX against RCC. The purpose of this study is to investigated the gene transcriptional alteration and investigate possible molecular mechanism and pathways implicated in the synergy of DAC and PTX against RCC.
cDNA microarray was performed and coupled with real-time PCR to identify critical genes in the synergistic mechanism of both agents against RCC cells. Various patterns of gene expression were observed by cluster analysis. IPA software was used to analyze possible biological pathways and to explore the inter-relationships between interesting network genes.
We found that lymphoid enhancer-binding factor 1 (LEF1), transforming growth factor β-induced (TGFBI), C-X-C motif ligand 5 (CXCL5) and myelocytomatosis viral related oncogene (c-myc) may play a pivotal role in the synergy of DAC and PTX. The PI3K/Akt pathway and other pathways associated with cyclins, DNA replication and cell cycle/mitotic regulation were also associated with the synergy of DAC and PTX against RCC.
The activation of PI3K/Akt-LEF1/β-catenin pathway could be suppressed synergistically by two agents and that PI3K/Akt-LEF1/β-catenin pathway is participated in the synergy of two agents.
Additional file 1: DAC (A) and PTX (B, C) each caused dosage-dependent cell growth suppression of RCC cells. DAC (0.5 and 1 μM) increased the susceptibility of RCC cells to PTX (B for ACHN and C for NC 65 are shown). The combination of DAC and PTX caused synergistic growth suppression in all two RCC cell lines by isobolographic analysis (D). (DOC 332 KB)
Additional file 2: Table S1 The top 10 up/down-regulated genes in the three different conditions normalized by untreated control.(JPEG 730 KB)
Additional file 3: Table S2 The top 10 up/down-regulated synergy-related genes by DAC and PTX are shown in (A) and (B), and synergy-related pathways by DAC and/or PTX are shown in (C).(JPEG 734 KB)
Mulders P, Brouwers A, Hulsbergen-van Der Kaa C, Van Lin E, Osanto S, De Mulder PH: ‘Guideline Renal cell carcinoma’. Ned Tijdschr Geneeskd. 2008, 152: 376-380. PubMed
Hartmann JT, Bokemeyer C: Chemotherapy for renal cell carcinoma. Anticancer Res. 1999, 19: 1541-1543. PubMed
Ravaud A, Audhuy B, Gomez F, Escudier B, Chevreau C, Douillard JY, Caty A, Geoffrois L, Ferrero JM, Linassier C, Drevon M, Négrier S: Subcutaneous interleukin-2, interferonα-2a, and continuous infusion of fluorouracil in metastatic renal cell carcinoma: a multicenter Phase II trial. J Clin Oncol. 1998, 16: 2728-2732. PubMed
Atzpodien J, Kirchner H, Hanninen E, Deckert M, Fenner M, Poliwoda H: Interleukin-2 in combination with interferon-αand 5-fluorouracil for metastatic renal cell carcinoma. Eur J Cancer. 1993, Suppl 5: S6-8. CrossRef
Chalitchagorn K, Shuangshoti S, Hourpai N, Kongruttanachok N, Tangkijvanich P, Thong-ngam D, Voravud N, Sriuranpong V, Mutirangura A: Distinctive pattern of LINE-1 methylation level in normal tissues and the association with carcinogenesis. Oncogene. 2004, 23: 8841-8846. 10.1038/sj.onc.1208137. CrossRefPubMed
Karpf AR, Moore BC, Ririe TO, Jones DA: Activation of the p53 DNA damage response pathway after inhibition of DNA methyltransferase by 5-aza-2′-deoxycytidine. Mol Pharmacol. 2001, 59: 751-757. PubMed
Kaufmann WK, Paules RS: DNA damage and cell cycle checkpoints. FASEB J. 1996, 10: 238-247. PubMed
Hattori K, Akaza H: New combination chemotherapy in urological cancers. Gan To Kagaku Ryoho. 2000, 27: 382-387. PubMed
Kaminski R, Kozar K, Niderla J, Grzela T, Wilczynski G, Skierski JS, Koronkiewicz M, Jakobisiak M, Golab J: Demethylating agent 5-aza-2′-deoxycytidine enhances expression of TNFRI and promotes TNF-mediated apoptosis in vitro and in vivo. Oncol Rep. 2004, 12: 509-516. PubMed
Alleman WG, Tabios RL, Chandramouli GV, Aprelikova ON, Torres-Cabala C, Mendoza A, Rogers C, Sopko NA, Linehan WM, Vasselli JR: The in vitro and in vivo effects of re-expressing methylated von Hippel-Lindau tumor suppressor gene in clear cell renal carcinoma with 5-aza-2′-deoxycytidine. Clin Cancer Res. 2004, 10: 7011-7021. 10.1158/1078-0432.CCR-04-0516. CrossRefPubMed
Shi H, Wei SH, Leu YW, Rahmatpanah F, Liu JC, Yan PS, Nephew KP, Huang TH: Triple analysis of the cancer epigenome: an integrated microarray system for assessing gene expression, DNA methylation, and histone acetylation. Cancer Res. 2003, 63: 2164-2171. PubMed
Shang D, Liu Y, Xu X, Han T, Tian Y: 5-aza-2′-deoxycytidine enhances susceptibility of renal cell carcinoma to paclitaxel by decreasing LEF1/phospho-β-catenin expression. Cancer Letter. 2011, 311: 230-236. 10.1016/j.canlet.2011.08.012. CrossRef
Kim JE, Jeong HW, Nam JO, Lee BH, Choi JY, Park RW, Park JY, Kim IS: Identification of motifs in the fasciclin domains of the transforming growth factor-beta-induced matrix protein betaig-h3 that interact with the alphavbeta5 integrin. J Biol Chem. 2002, 277: 46159-46165. 10.1074/jbc.M207055200. CrossRefPubMed
Skonier J, Neubauer M, Madisen L, Bennett K, Plowman GD, Purchio AF: cDNA cloning and sequence analysis of beta ig-h3, a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor-beta. DNA Cell Biol. 1992, 11: 511-522. 10.1089/dna.1992.11.511. CrossRefPubMed
Ivanov SV, Ivanova AV, Salnikow K, Timofeeva O, Subramaniam M, Lerman MI: Two novel VHL targets, TGFBI (BIGH3) and its transactivator KLF10, are up-regulated in renal clear cell carcinoma and other tumors. Biochem Biophys Res Commun. 2008, 370: 536-540. 10.1016/j.bbrc.2008.03.066. PubMedCentralCrossRefPubMed
Shang D, Liu Y, Yang P, Chen Y, Tian Y: TGFBI-promoted adhesion, migration and invasion of human renal cell carcinoma depends on inactivation of von Hippel-Lindau tumor suppressor. Urology. 2012, 79: 966.e1-966.e7. 10.1016/j.urology.2011.12.011. CrossRef
Wong YF, Cheung TH, Lo KW, Yim SF, Siu NS, Chan SC, Ho TW, Wong KW, Yu MY, Wang VW, Li C, Gardner GJ, Bonome T, Johnson WB, Smith DI, Chung TK, Birrer MJ: Identification of molecular markers and signaling pathway in endometrial cancer in Hong Kong Chinese women by genome-wide gene expression profiling. Oncogene. 2007, 26: 1971-1982. 10.1038/sj.onc.1209986. CrossRefPubMed
Napoli C, Lerman LO, de Nigris F, Sica V: c-myc oncoprotein: a dual pathogenic role in neoplasia and cardiovascular diseases?. Neoplasia. 2002, 3: 185-190. CrossRef
Coller HA, Grandori C, Tamayo P, Colbert T, Lander ES, Eisenman RN, Golub TR: Expression analysis with oligonucleotide microarrays reveals that MYC regulates genes involved in growth, cell cycle, signaling, and adhesion. Proc Natl Acad Sci. 2000, 97: 3260-3265. 10.1073/pnas.97.7.3260. PubMedCentralCrossRefPubMed
Bova GS, Carter BS, Bussemakers MJ, Emi M, Fujiwara Y, Kyprianou N, Jacobs SC, Robinson JC, Epstein JI, Walsh PC: Homozygous deletion and frequent allelic loss of chromosome 8p22 loci in human prostate cancer. Cancer Res. 1993, 53: 3869-3873. PubMed
Macoska JA, Trybus TM, Benson PD, Sakr WA, Grignon DJ, Wojno KD, Pietruk T, Powell IJ: Evidence for three tumor suppressor gene loci on chromosome 8p in human prostate cancer. Cancer Res. 1995, 55: 5390-5395. PubMed
Furge KA, Chen J, Koeman J, Swiatek P, Dykema K, Lucin K, Kahnoski R, Yang XJ, Teh BT: Detection of DNA copy number changes and oncogenic signaling abnormalities from gene expression data reveals MYC activation in high-grade papillary renal cell carcinoma. Cancer Res. 2007, 67: 3171-3176. 10.1158/0008-5472.CAN-06-4571. CrossRefPubMed
Schraml P, Kononen J, Bubendorf L, Moch H, Bissig H, Nocito A, Mihatsch MJ, Kallioniemi OP, Sauter G: Tissue microarrays for gene amplification surveys in many different tumor types. Clin Cancer Res. 1999, 5: 1966-1975. PubMed
Manna P, Jain SK: Hydrogen sulfide and L-cysteine increase phosphatidylinositol 3,4,5-trisphosphate (PIP3) and glucose utilization by inhibiting phosphatase and tensin homolog (PTEN) protein and activating phosphoinositide 3-kinase (PI3K)/serine/threonine protein kinase (AKT)/protein kinase Cζ/λ (PKCζ/λ) in 3T3l1 adipocytes. J Biol Chem. 2011, 286: 39848-39859. 10.1074/jbc.M111.270884. PubMedCentralCrossRefPubMed
- Gene expression profiling of the synergy of 5-aza-2′-deoxycytidine and paclitaxel against renal cell carcinoma
- BioMed Central
Neu im Fachgebiet Chirurgie
Mail Icon II