Background
Results
Global transcript profiling reveals novel PRL-modulated genes in human T47D breast cancer xenotransplants
Unigene | Symbol | Gene descriptor | Fold change | |
---|---|---|---|---|
1 | Hs.89626 | PTHrP | parathyroid hormone-like hormone | 11.9 |
2 | Hs.473539 | BACH1 | BTB and CNC homology 1, basic leucine zipper transcription factor 1 | 5.2 |
3 | Hs.46468 | CCR6 | chemokine (C-C motif) receptor 6 | 4.9 |
4 | Hs.314676 | ITCH | itchy homolog E3 ubiquitin protein ligase | 4.2 |
5 | Hs.150744 | INVS | inversin | 3.8 |
6 | Hs.387222 | NEK6 | NIMA (never in mitosis gene a)-related kinase 6 | 3.8 |
7 | Hs.76095 | IER3 | immediate early response 3 | 3.4 |
8 | Hs.121520 | AMIGO2 | amphoterin induced gene 2 | 3.3 |
9 | Hs.115263 | EREG | epiregulin | 3.2 |
10 | Hs.436186 | ERAP1 | type 1 tumor necrosis factor receptor shedding aminopeptidase regulator | 3.0 |
11 | Hs.99037 | CTEN | C-terminal tensin-like | 3.0 |
12 | Hs.252855 | MFI2 | antigen p97 (melanoma associated) | 2.9 |
13 | Hs.439658 | MGC4796 | Ser/Thr-like kinase | 2.8 |
14 | Hs.417962 | DUSP4 | dual specificity phosphatase 4 | 2.7 |
15 | Hs.8257 | CISH | cytokine inducible SH2-containing protein | 2.6 |
16 | Hs.660427 | PAR5 | Prader-Willi/Angelman syndrome-5 | 2.6 |
17 | Hs.279887 | AIPL1 | aryl hydrocarbon receptor interacting protein-like 1 | 2.6 |
18 | Hs.145807 | TMC5 | transmembrane channel-like 5 | 2.5 |
19 | Hs.512708 | TGM2 | transglutaminase 2 | 2.4 |
20 | Hs.270833 | AREG | amphiregulin (schwannoma-derived growth factor) | 2.4 |
21 | Hs.170623 | FGD6 | FYVE, RhoGEF and PH domain containing 6 | 2.3 |
22 | Hs.418138 | FN1 | fibronectin 1 | 2.3 |
23 | Hs.102541 | NTN4 | netrin 4 | 2.2 |
24 | Hs.354906 | RAB39 | RAB39, member RAS oncogene family | 2.2 |
25 | Hs.443906 | EGLN3 | egl nine homolog 3 (C. elegans) | 2.2 |
26 | Hs.149156 | GLDC | glycine dehydrogenase | 2.2 |
27 | Hs.1145 | WT1 | Wilms tumor 1 | 2.1 |
28 | Hs.25220 | LARGE | like-glycosyltransferase | 2.1 |
29 | Hs.282557 | CP | ceruloplasmin (ferroxidase) | 2.0 |
30 | Hs.78909 | ZFP36L2 | zinc finger protein 36, C3H type-like 2 | 2.0 |
31 | Hs.413297 | RGS16 | regulator of G-protein signalling 16 | 2.0 |
32 | Hs.182454 | NYREN18 | NEDD8 ultimate buster-1 | 2.0 |
33 | Hs.96125 | RCP | Rab coupling protein | 2.0 |
34 | Hs.308028 | TMEM17 | transmembrane protein 17 | 2.0 |
35 | Hs.21894 | PPM1H | protein phosphatase 1H (PP2C domain containing) | 2.0 |
36 | Hs.240395 | KCNK6 | potassium channel, subfamily K, member 6 | 1.9 |
37 | Hs.36563 | B7-H4 | immune costimulatory protein B7-H4 | 1.9 |
38 | Hs.27345 | RNGTT | RNA guanylyltransferase and 5’-phosphatase | 1.9 |
39 | Hs.144287 | HEY2 | hairy/enhancer-of-split related with YRPW motif 2 | 1.8 |
40 | Hs.269857 | HRB2 | HIV-1 rev binding protein 2 | 1.8 |
41 | Hs.387871 | TNFSF10 | tumor necrosis factor (ligand) superfamily, member 10 | 1.8 |
42 | Hs.80409 | GADD45A | growth arrest and DNA-damage-inducible, alpha | 1.8 |
43 | Hs.134742 | FAM20C | family with sequence similarity 20, member C | 1.8 |
44 | Hs.274701 | TK2 | thymidine kinase 2, mitochondrial | 1.8 |
45 | Hs.55610 | SLC30A1 | solute carrier family 30 (zinc transporter), member 1 | 1.8 |
46 | Hs.82173 | TIEG | TGFB inducible early growth response | 1.8 |
47 | Hs.404081 | FGFR2 | fibroblast growth factor receptor 2 | 1.8 |
48 | Hs.31218 | SCAMP1 | secretory carrier membrane protein 1 | 1.8 |
49 | Hs.350470 | TFF1 | trefoil factor 1 | 1.8 |
50 | Hs.95655 | SECTM1 | secreted and transmembrane 1 | 1.8 |
51 | Hs.902 | NF2 | neurofibromin 2 | 1.7 |
52 | Hs.310640 | T2BP | TRAF2 binding protein | 1.7 |
53 | Hs.252550 | TNIK | TRAF2 and NCK interacting kinase | 1.7 |
54 | Hs.6838 | ARHE | ras homolog gene family, member E | 1.7 |
55 | Hs.418062 | B3GALT3 | betaGlcNAc beta 1,3-galactosyltransferase, polypeptide 3 | 1.7 |
56 | Hs.270411 | PLEKHC1 | pleckstrin homology domain containing, family C member 1 | 1.7 |
57 | Hs.647388 | ARHGDIG | Rho GDP dissociation inhibitor (GDI) gamma | 1.7 |
58 | Hs.202453 | MYC | v-myc myelocytomatosis viral oncogene homolog (avian) | 1.6 |
59 | Hs.110488 | CHSY1 | carbohydrate (chondroitin) synthase 1 | 1.6 |
60 | Hs.9795 | ACOX2 | acyl-Coenzyme A oxidase 2, branched chain | 1.6 |
61 | Hs.158357 | UNC5CL | unc-5 homolog C (C. elegans)-like | 1.6 |
62 | Hs.441972 | IFNT1 | interferon tau-1 | 1.6 |
63 | Hs.221889 | CSDA | cold shock domain protein A | 1.6 |
64 | Hs.333503 | RNF38 | ring finger protein 38 | 1.6 |
65 | Hs.203581 | DDX54 | DEAD (Asp-Glu-Ala-Asp) box polypeptide 54 | 1.6 |
66 | Hs.345226 | ZNF563 | zinc finger protein 563 | 1.6 |
67 | Hs.30991 | ANKRD6 | ankyrin repeat domain 6 | 1.6 |
68 | Hs.4113 | AHCYL1 | S-adenosylhomocysteine hydrolase-like 1 | 1.6 |
69 | Hs.416077 | SEMA4B | sema domain | 1.6 |
70 | Hs.7378 | PHLDB2 | pleckstrin homology-like domain, family B, member 2 | 1.6 |
71 | Hs.369063 | ZIC2 | Zic family member 2 (odd-paired homolog, Drosophila) | 1.6 |
72 | Hs.515284 | ZNF505 | zinc finger protein 505 | 1.6 |
73 | Hs.426511 | MIPOL1 | mirror-image polydactyly 1 | 1.6 |
74 | Hs.108966 | PIP5K2A | phosphatidylinositol-4-phosphate 5-kinase, type II, alpha | 1.6 |
75 | Hs.432607 | PSMB2 | proteasome (prosome, macropain) subunit, beta type, 2 | 1.6 |
Unigene | Symbol | Gene descriptor | Fold Change | |
---|---|---|---|---|
1 | Hs.275464 | KLK10 | kallikrein 10 | −3.3 |
2 | Hs.307030 | KRTAP1-5 | keratin associated protein 1-5 | −3.3 |
3 | Hs.282233 | MLLT6 | myeloid/lymphoid leukemia translocated to, 6 | −3.2 |
4 | Hs.78518 | NPR2 | natriuretic peptide receptor B/guanylate cyclase B | −2.8 |
5 | Hs.155024 | BCL6 | B-cell CLL/lymphoma 6 | −2.8 |
6 | Hs.90800 | MMP16 | matrix metalloproteinase 16 (membrane-inserted) | −2.5 |
7 | Hs.87539 | ALDH3B2 | aldehyde dehydrogenase 3 family, member B2 | −2.4 |
8 | Hs.144906 | METAP2 | methionyl aminopeptidase 2 | −2.3 |
9 | Hs.440455 | ALAS2 | aminolevulinate, delta-, synthase 2 | −2.2 |
10 | Hs.266175 | PAG | phosphoprotein associated with glycosphingolipid-enriched microdomains | −2.2 |
11 | Hs.435947 | RBM15 | RNA binding motif protein 15 | −2.2 |
12 | Hs.233325 | HFE | hemochromatosis | −2.1 |
13 | Hs.443012 | SEMA6A | sema domain,transmembrane domain(TM),and cytoplasmic domain,(semaphorin)6A | −2.0 |
14 | Hs.357901 | SOX4 | SRY (sex determining region Y)-box 4 | −2.0 |
15 | Hs.415048 | FLT4 | fms-related tyrosine kinase 4 | −2.0 |
16 | Hs.380833 | IGFBP5 | insulin-like growth factor binding protein 5 | −1.9 |
17 | Hs.444881 | CRAMP1L | Crm, cramped-like (Drosophila) | −1.9 |
18 | Hs.398124 | DNAH5 | dynein, axonemal, heavy polypeptide 5 | −1.9 |
19 | Hs.79025 | SNRK | SNF-1 related kinase | −1.9 |
20 | Hs.432121 | PRDX2 | peroxiredoxin 2 | −1.9 |
21 | Hs.22370 | NEXN | nexilin (F actin binding protein) | −1.9 |
22 | Hs.144914 | GNMT | glycine N-methyltransferase | −1.9 |
23 | Hs.21446 | CENTB5 | centaurin, beta 5 | −1.9 |
24 | Hs.58103 | AKAP9 | A kinase (PRKA) anchor protein (yotiao) 9 | −1.8 |
25 | Hs.387385 | SMURF2 | E3 ubiquitin ligase SMURF2 | −1.8 |
26 | Hs.324470 | ADD3 | adducin 3 (gamma) | −1.8 |
27 | Hs.348387 | GSTM4 | glutathione S-transferase M4 | −1.8 |
28 | Hs.58419 | TARSH | target of Nesh-SH3 | −1.7 |
29 | Hs.174051 | SNRP70 | small nuclear ribonucleoprotein 70 kDa polypeptide (RNP antigen) | −1.7 |
30 | Hs.211601 | MAP3K12 | mitogen-activated protein kinase kinase kinase 12 | −1.7 |
31 | Hs.23964 | SAP18 | sin3-associated polypeptide, 18 kDa | −1.7 |
32 | Hs.380929 | LDHD | lactate dehydrogenase D | −1.7 |
33 | Hs.390568 | ZNF585A | zinc finger protein 585A | −1.7 |
34 | Hs.241305 | TRIM16 | tripartite motif-containing 16 | −1.7 |
35 | Hs.403933 | FBXO32 | F-box only protein 32 | −1.7 |
36 | Hs.434756 | AP2E | adaptor-related protein complex 2, epsilon subunit | −1.7 |
37 | Hs.173894 | CSF1 | colony stimulating factor 1 (macrophage) | −1.6 |
38 | Hs.104555 | NPFF | neuropeptide FF-amide peptide precursor | −1.6 |
39 | Hs.307015 | KRTAP4-14 | keratin associated protein 4-14 | −1.6 |
40 | Hs.301961 | GSTM1 | glutathione S-transferase M1 | −1.6 |
41 | Hs.307915 | ABCC4 | ATP-binding cassette, sub-family C (CFTR/MRP), member 4 | −1.6 |
42 | Hs.512000 | GP1BB | glycoprotein Ib (platelet), beta polypeptide | −1.6 |
43 | Hs.446297 | ZNF498 | zinc finger protein 498 | −1.6 |
44 | Hs.120396 | FRMD4 | FERM domain containing 4 | −1.6 |
45 | Hs.222901 | GRIK4 | glutamate receptor, ionotropic, kainate 4 | −1.6 |
46 | Hs.464896 | ZNF397 | zinc finger protein 397 | −1.6 |
47 | Hs.16232 | CNKSR1 | connector enhancer of kinase suppressor of Ras 1 | −1.6 |
48 | Hs.255526 | DTNA | dystrobrevin, alpha | −1.6 |
49 | Hs.107203 | PLAC2 | placenta-specific 2 | −1.6 |
50 | Hs.9029 | KRT23 | keratin 23 (histone deacetylase inducible) | −1.6 |
51 | Hs.91753 | SMPD3 | sphingomyelin phosphodiesterase 3, neutral membrane (neutral sphingomyelinase II) | −1.6 |
52 | Hs.331555 | SPINK5 | serine protease inhibitor, Kazal type 5 | −1.6 |
53 | Hs.391858 | TIA1 | TIA1 cytotoxic granule-associated RNA binding protein | −1.6 |
54 | Hs.109122 | MPP5 | membrane protein, palmitoylated 5 (MAGUK p55 subfamily member 5) | −1.6 |
55 | Hs.445072 | ARGBP2 | Arg/Abl-interacting protein ArgBP2 | −1.6 |
Unigene | Symbol | Gene descriptor | Fold Change | |
---|---|---|---|---|
1 | Hs.89626 | PTHrP | parathyroid hormone-like hormone | 11.9 |
2 | Hs.46468 | CCR6 | chemokine (C-C motif) receptor 6 | 4.9 |
3 | Hs.76095 | IER3 | immediate early response 3 | 3.4 |
4 | Hs.121520 | AMIGO2 | amphoterin induced gene 2 | 3.3 |
5 | Hs.115263 | EREG | epiregulin | 3.2 |
6 | Hs.436186 | ERAP1 | type 1 tumor necrosis factor receptor shedding aminopeptidase regulator | 3.0 |
7 | Hs.417962 | DUSP4 | dual specificity phosphatase 4 | 2.7 |
8 | Hs.8257 | CISH | cytokine inducible SH2-containing protein | 2.6 |
9 | Hs.145807 | TMC5 | transmembrane channel-like 5 | 2.5 |
10 | Hs.270833 | AREG | amphiregulin (schwannoma-derived growth factor) | 2.4 |
11 | Hs.418138 | FN1 | fibronectin 1 | 2.3 |
12 | Hs.102541 | NTN4 | netrin 4 | 2.2 |
13 | Hs.1145 | WT1 | Wilms tumor 1 | 2.1 |
14 | Hs.80409 | GADD45A | growth arrest and DNA-damage-inducible, alpha | 1.8 |
15 | Hs.404081 | FGFR2 | fibroblast growth factor receptor 2 | 1.8 |
16 | Hs.155024 | BCL6 | B-cell CLL/lymphoma 6 | −2.8 |
17 | Hs.357901 | SOX4 | SRY (sex determining region Y)-box 4 | −2.0 |
18 | Hs.415048 | FLT4 | fms-related tyrosine kinase 4 | −2.0 |
qRT-PCR validation of 18 candidate prolactin modulated genes
Stat5 regulates novel PRL-modulated transcripts
Jak2 and Jak1 are critical for PRL gene regulation in T47D breast cancer cells
Synergistic enhancement by E2 of PRL-induced proliferation and select PRL-modulated genes
PTHrP protein levels correlate with levels of pYStat5 in human breast cancer tissues
Gene ontology (GO) terms are enriched in differentiation markers
GO Term | Description | P value | FDR |
---|---|---|---|
GO:0030005 | cellular di-, tri-valent inorganic cation homeostasis | 0.002 | 2.452 |
GO:0042592 | homeostatic process | 0.002 | 2.848 |
GO:0055066 | di-, tri-valent inorganic cation homeostasis | 0.002 | 3.079 |
GO:0030003 | cellular cation homeostasis | 0.003 | 4.015 |
GO:0055080 | cation homeostasis | 0.004 | 6.550 |
GO:0006879 | cellular iron ion homeostasis | 0.006 | 8.210 |
GO:0043123 | positive regulation of I-kappaB kinase/NF-kappaB cascade | 0.006 | 9.136 |
GO:0060249 | anatomical structure homeostasis | 0.008 | 10.970 |
GO:0055072 | iron ion homeostasis | 0.008 | 11.045 |
GO:0019725 | cellular homeostasis | 0.008 | 11.791 |
GO:0043122 | regulation of I-kappaB kinase/NF-kappaB cascade | 0.008 | 11.809 |
GO:0048514 | blood vessel morphogenesis | 0.009 | 12.950 |
GO:0051052 | regulation of DNA metabolic process | 0.010 | 13.580 |
GO:0042127 | regulation of cell proliferation | 0.010 | 13.992 |
GO:0043066 | negative regulation of apoptosis | 0.011 | 15.532 |
GO:0043069 | negative regulation of programmed cell death | 0.012 | 16.371 |
GO:0060548 | negative regulation of cell death | 0.012 | 16.542 |
GO:0048878 | chemical homeostasis | 0.013 | 17.642 |
GO:0006873 | cellular ion homeostasis | 0.014 | 18.854 |
GO:0055082 | cellular chemical homeostasis | 0.014 | 19.975 |
GO:0001568 | blood vessel development | 0.015 | 20.556 |
GO:0010627 | regulation of protein kinase cascade | 0.015 | 20.807 |
GO:0001944 | vasculature development | 0.016 | 22.079 |
Prolactin-upregulated gene signature is associated with prolonged time to relapse and metastasis
Gene Symbol | Correlation | Gene symbol | Correlation |
---|---|---|---|
with signature average | with signature average | ||
AREG | 0.6 | TNIK | 0.25 |
DUSP4 | 0.51 | PPM1H | 0.17 |
PTHLH | 0.46 | PAR5 | 0.15 |
EREG | 0.45 | INVS | 0.14 |
SCAMP1 | 0.43 | CSDA | 0.13 |
TMC5 | 0.42 | MYC | 0.12 |
GADD45A | 0.42 | WT1 | 0.09 |
CHSY1 | 0.41 | CISH | 0.08 |
TFF1 | 0.4 | CP | 0.07 |
FGD6 | 0.39 | RGS16 | 0.07 |
BACH1 | 0.38 | NF2 | 0.07 |
AHCYL1 | 0.37 | FGFR2 | 0.04 |
EGLN3 | 0.34 | RNGTT | 0.03 |
AMIGO2 | 0.33 | FN1 | 0.02 |
TNFSF10 | 0.33 | ITCH | −0.01 |
IER3 | 0.33 | LARGE | −0.11 |
ANKRD6 | 0.32 | PSMB2 | −0.12 |
ACOX2 | 0.31 | SECTM1 | −0.15 |
SLC30A1 | 0.31 | GLDC | −0.2 |
ERAP1 | 0.31 | AIPL1 | −0.21 |
RNF38 | 0.3 | TGM2 | −0.22 |
ZFP36L2 | 0.3 | DDX54 | −0.23 |
CCR6 | 0.28 | MFI2 | −0.27 |
HEY2 | 0.25 | ARHGDIG | −0.27 |
TK2 | 0.25 |
Discussion
Gene | Fold Change (microarray) | in vivo Regulation | in vitro Regulation | Stat5 Regulation | Jak2 Regulation | Jak1 Regulation | E2 Regulation |
---|---|---|---|---|---|---|---|
PTHrP | 11.9 | Y | Y | Y | Y | Y | Y |
CCR6 | 4.9 | Y | N | Y | Y | Y | Y |
IER3 | 3.4 | Y | Y | Y | Y | Y | Y |
AMIGO2 | 3.3 | Y | N | NA | NA | NA | NA |
EREG | 3.2 | Y | Y | Y | Y | Y | Y |
ERAP1 | 3.0 | N | N | NA | NA | NA | NA |
DUSP4 | 2.7 | Y | N | NA | NA | NA | NA |
CISH | 2.6 | Y | Y | Y | Y | Y | Y |
TMC5 | 2.5 | Y | Y | Y | Y | Y | N |
AREG | 2.4 | Y | Y | Y | Y | Y | Y |
FN1 | 2.3 | Y | Y | Y | Y | Y | Y |
NTN4 | 2.2 | Y | Y | Y | Y | Y | N |
WT1 | 2.1 | Y | Y | Y | Y | Y | Y |
GADD45A | 1.8 | N | N | NA | NA | NA | NA |
FGFR2 | 1.8 | N | N | NA | NA | NA | NA |
BCL6 | −2.8 | Y | Y | Y | Y | N | N |
SOX4 | −2.0 | Y | Y | Y | NA | NA | N |
FLT4 | −2.0 | Y | Y | Y | NA | NA | N |
Conclusions
Materials and methods
Tissue culture
Xenotransplants
Microarray
Quantitative reverse transcription polymerase chain reaction
Gene | Forward primer | Reverse primer |
---|---|---|
PTHrP | GTTCCTGGTGAGCTACGCG | CTTGGATGGACTTCCCCTTG |
CCR6 | TGCTACCGCTGCCTGTGAGC | AAAATAATCTTCACTGGAGTCG |
IER3 | CGTCCTCGAGCCCTTTAATCT | AGGTCCAGAGCGTAGTCCGA |
AMIGO2 | CCGGTGTCTTTTCCACCG | GAGCCCACGAGGCTCC |
EREG | GCTCTGCCTGGGTTTCCATC | CCACACGTGGATTGTCTTCTGTC |
ERAP1 | GCCATTCTAGCTGCAGTGGG | CAACTGTGTACGGGAGCCC |
DUSP4 | TACAAGTGCATCCCAGTGGA | CCCGTTTCTTCATCATCAGG |
CISH | CTGCTGTGCATAGCCAAGAC | GTGCCTTCTGGCATCTTCTG |
TMC5 | TATCCTTCAGCTCAATTGCTG | AGAGGACGCTGGTTCCAAAC |
AREG | GGTGGTGCTGTCGCTCTTG | TCAGCACTGTGGTCCCCAG |
FN1 | TTCTACTCCTGCACACAGAAG | CCCTCAGAAGTGCAATCAGTG |
NTN4 | CATGGTGGGATACTGGGGC | TCAGGAACTTCATGATACCAGTC |
WT1 | GAGAGCCAGCCCGCTATTC | CATGGGATCCTCATGCTTG |
GADD45A | TCAGCGCACGATCACTGTC | CCAGCAGGCACAACACCAC |
FGFR2 | CTCACTCTCACAACCAATGAGG | AGGAAGGCATGGTTCGTAAG |
BCL6 | CTGCAGATGGAGCATGTTGT | TCTTCACGAGGAGGCTTGAT |
SOX4 | GTGAGCGAGATGATCTCGGG | CAGGTTGGAGATGCTGGACTC |
FLT4 | CAGGATGAAGACATTTGAGG | AAGAAAATGCTGACGTAT |
GAPDH | AATCCATCACCATCTTCCA | TGGACTCCACGACGTACTCA |
ERAP1 | GCCATTCTAGCTGCAGTGGG | CAACTGTGTACGGGAGCCC |