684
N. Gokan et al. / Biochemical Pharmacology 70 (2005) 676–685
[7] Massento MS, Morris HR, Taylor GW, Johnson SJ, Skapski AC, Kay
RR. Differentiation-inducing factor from the slime mould Dictyoste-
lium discoideum and its analogues. Biochem J 1988;256:23–8.
[8] Morris HR, Masento MS, Taylor GW, Jermyn KA, Kay RR. Structure
elucidation of two differentiation inducing factors (DIF-2 and DIF-3)
from the cellular slime mould Dictyostelium discoideum. Biochem J
1988;249:903–6.
K562 cells (Figs. 1 and 4). Analyses of many other DIF
derivatives may give us hints for identifying the receptor(s)
of DIF-1 in D. discoideum in the future.
4.2. Anti-proliferative activity of DIF analogues in
K562 cells
[9] Kay RR, Flatman P, Thompson CRL. DIF signalling and cell fate.
Semin Cell Dev Biol 1999;10:577–85.
[10] Kubohara Y, Okamoto K. Cytoplasmic Ca2+ and H+ concentrations
determine cell fate in Dictyostelium discoideum. Federation Am Soc
Exp Biol J 1994;8:869–74.
As already mentioned, DIFs possess anti-tumor activ-
ities in several cell lines [13–17,20,21], and the order of
potency for anti-proliferative activity was established as
DIF-3 > DIF-1 > DIF-2 (designated DIF-1(ꢀ1) in this
study) in K562 cells [17]. Here, we have added interesting
profiles of the many newly synthesized DIF analogues. Of
the present findings, it is noteworthy that Bu-DIF-3 and
CP-DIF-3 possess strong anti-proliferative activity
(Fig. 5C). This would be valuable information in the
development of more effective and useful derivatives of
DIF-3 and indicates that the pharmacological activity of
some other derivatives having modifications at the position
of the methoxy group in DIF-3 should be examined in the
future.
[11] Schaap P, Nebl T, Fisher PR. A slow sustained increase in cytosolic
Ca2+ levels mediates stalk gene induction by differentiation inducing
factor in Dictyostelium. Eur Mol Biol Organ J 1996;15:5177–83.
[12] Azhar M, Kennady PK, Pande G, Nanjundiah V. Stimulation by DIF
causes an increase of intracellular Ca2+ in Dictyostelium discoideum.
Exp Cell Res 1997;230:403–6.
[13] Asahi K, Sakurai A, Takahashi N, Kubohara Y, Okamoto K, Tanaka Y.
DIF-1, morphogen of Dictyostelium discoideum, induces the erythroid
differentiation in murine and human leukemia cells. Biochem Biophys
Res Commun 1995;208:1036–9.
[14] Kubohara Y, Saito Y, Tatemoto K. Differentiation-inducing factor of
D. discoideum raises intracellular calcium concentration and sup-
presses cell growth in rat pancreatic AR42J cells. Federation Eur
Biochem Soc Lett 1995;359:119–22.
Recently, we reported that PDE1 is a pharmacological
and specific target of DIFs [23]. However, some of the
known actions of DIFs in mammalian cells do not seem to
be exerted via PDE1 inhibition by DIFs. For example, a
rise in [Ca2+]i by DIFs [14,16,17] may not be explained by
PDE1 inhibition by them, suggesting that there should
be some other target(s) of DIFs in mammalian cells. The
present findings would give us hints for identifying the
pharmacological target(s) of DIFs in mammalian cells and
also for developing novel anti-cancer drugs.
[15] Kubohara Y, Kimura C, Tatemoto K. Putative morphogen, DIF, of
Dictyostelium discoideum induces apoptosis in rat pancreatic AR42J
cells. Dev Growth Differ 1995;37:711–6.
[16] Kubohara Y. DIF-1, putative morphogen of D. discoideum, suppresses
cell growth and promotes retinoic acid-induced cell differentiation in
HL-60. Biochem Biophys Res Commun 1997;236:418–22.
[17] Kubohara Y. Effects of differentiation-inducing factors (DIFs) of
Dictyostelium discoideum on the human leukemia K562 cells: DIF-3
isthemostpotentanti-leukemicagent.EurJPharmacol1999;381:57–62.
[18] Kubohara Y, Hosaka K. The putative morphogen, DIF-1, of Dictyos-
telium discoideum activates Akt/PKB in human leukemia K562 cells.
Biochem Biophys Res Commun 1999;263:790–6.
[19] Miwa Y, Sasaguri T, Kosaka C, Taba Y, Ishida A, Abumiya T, et al.
DIF-1, a morphogen of Dictyostelium, induces G1 arrest and differ-
entiation of vascular smooth muscle cells. Circ Res 2000;86:68–75.
[20] Kanai M, Konda Y, Nakajima T, Izumi Y, Nanakin A, Kanda N, et al.
Differentiation-inducing factor-1 (DIF-1) inhibits STAT3 activity
involved in gastric cancer cell proliferation via MEK-ERK dependent
pathway. Oncogene 2003;22:548–54.
Acknowledgement
This work was supported in parts by grants from the
Ministry of Education, Science, and Culture of Japan (Y.K.
and K.H.).
[21] Takahashi-Yanaga F, Taba Y, Miwa Y, Kubohara Y, Watanabe Y,
Hirata M, et al. Dictyostelium differentiation-inducing factor-3 acti-
vates glycogen synthase kinase-3b and degrades cyclin D1 in mam-
malian cells. J Biol Chem 2003;278:9663–70.
References
[22] Akaishi E, Narita T, Kawai S, Miwa Y, Sasaguri T, Hosaka K, et al.
Differentiation-inducing factor-1-induced growth arrest of K562 leu-
kemia cells involves the reduction of ERK1/2 activity. Eur J Pharma-
col 2004;485:21–9.
[1] Konijn TM, Van de Meene JGC, Bonner JT, Barkley DS. The acrasin
activity of adenosine-30,50-cyclic phosphate. Proc Natl Acad Sci USA
1967;58:1152–4.
[2] Darmon M, Brachet P, Pereira da Silva LH. Chemotactic signals
induce cell differentiation in Dictyostelium discoideum. Proc Natl
Acad Sci USA 1975;72:3163–6.
[23] Shimizu K, Murata T, Tagawa T, Takahashi K, Ishikawa R, Abe Y,
et al. Calmodulin-dependent cyclic nucleotide phosphodiesterase
(PDE1) is a pharmacological target of differentiation-inducing fac-
tor-1, an anti-tumor agent isolated from Dictyostelium. Cancer Res
2004;64:2568–71.
[3] Gerisch G, Fromm H, Husesgen A, Wick U. Control of cell-contact
sites by cyclic AMP pulses in differentiating Dictyostelium cells.
Nature 1975;255:547–9.
[24] Grove JF, Jeffs PW, Rustidge DW. Griseofulvin. Part X. The orientation
ofsomederivativesof5-methoxyresorcinol. JChemSoc1956;1956–63.
[25] Kajigaeshi S, Kakinami T, Yamasaki H, Fujisaki S, Kondo M,
Okamoto T. Iodination of phenols by use of benzyltrimethylammo-
nium dichloroiodate. Chem Lett 1987;2109–12.
[4] Town CD, Gross JD, Kay RR. Cell differentiation without morpho-
genesis in Dictyostelium discoideum. Nature 1976;262:717–9.
[5] Morris HR, Taylor GW, Masento MS, Jermyn KA, Kay RR. Chemical
structure of the morphogen differentiation inducing factor from Dic-
tyostelium discoideum. Nature 1987;328:811–4.
[26] Barton DHR, Finet JP, Khamsi J, Pichon C. Copper catalysed
o-phenylation of phenols and enols by pentavalent organobismuth
compounds. Tetrahedron Lett 1986;27:3619–22.
[6] Kay RR, Berks M, Traynor D. Morphogen hunting in Dictyostelium
discoideum. Development 1989;Suppl.:81–90.