Dictyostelium differentiation‐inducing factor‐1 induces glucose transporter 1 translocation and promotes glucose uptake in mammalian cells

Abstract

The differentiation-inducing factor-1 (DIF-1) is a signal molecule that induces stalk cell formation in the cellular slime mold Dictyostelium discoideum, while DIF-1 and its analogs have been shown to possess antiproliferative activity in vitro in mammalian tumor cells. In the present study, we investigated the effects of DIF-1 and its analogs on normal (nontransformed) mammalian cells. Without affecting the cell morphology and cell number, DIF-1 at micromolar levels dose-dependently promoted the glucose uptake in confluent 3T3-L1 fibroblasts, which was not inhibited with wortmannin or LY294002 (inhibitors for phosphatidylinositol 3-kinase). DIF-1 affected neither the expression level of glucose transporter 1 nor the activities of four key enzymes involved in glucose metabolism, such as hexokinase, fluctose 6-phosphate kinase, pyruvate kinase, and glucose 6-phosphate dehydrogenase. Most importantly, stimulation with DIF-1 was found to induce the translocation of glucose transporter 1 from intracellular vesicles to the plasma membranes in the cells. In differentiated 3T3-L1 adipocytes, DIF-1 induced the translocation of glucose trasporter 1 (but not of glucose transporter 4) and promoted glucose uptake, which was not inhibited with wortmannin. These results indicate that DIF-1 induces glucose transporter 1 translocation and thereby promotes glucose uptake, at least in part, via a inhibitors for phosphatidylinositol 3-kinase ⁄ Akt-independent pathway in mammalian cells. Furthermore, analogs of DIF-1 that possess stronger antitumor activity than DIF-1 were less effective in promoting glucose consumption, suggesting that the mechanism of the action of DIF-1 for stimulating glucose uptake should be different from that for suppressing tumor cell growth.Abbreviations DIF-1, differentiation-inducing factor-1 [1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl)hexan-1-one]; DIF-3, differentiation-inducing factor-3 [1-(3-chloro-2,6-dihydroxy-4-methoxyphenyl)hexan-1-one]; DMEM-HG, DMEM containing a high concentration of glucose; DMEM-LG, DMEM containing a low concentration of glucose; EtOH, ethanol; GLUT, glucose transporter; LDM, low-density microsome; LY294002, 2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one; 8-MIBMX, 8-methoxymethyl-3-isobutyl-1-methylxanthine; 2-MIDIF-1, 2-methoxy isomer of DIF-1; PDE1, calmodulin-dependent cyclic nucleotide phosphodiesterase; PI3K, phosphatidylinositol 3-kinase; PM, plasma membrane; Tg, thapsigargin; THPH, 1-(2,4,6-trihydroxyphenyl)hexan-1-one.

Dictyostelium Differentiation-Inducing Factor-1 Binds to Mitochondrial Malate Dehydrogenase and Inhibits Its Activity

“…Alternatively, a recently reported finding that DIF-1, but not 2-MIDIF-1, was able to promote glucose uptake and consumption could explain the inhibition results (23). It is possible that DIF-1-treated cells deplete glucose in the medium during the culture period.…”

Biological Activities of Novel Derivatives of Differentiation-Inducing Factor 3 from <i>Dictyostelium discoideum</i>

“…D5796, Sigma-Aldrich) supplemented with 10% (v/v) fetal bovine serum (FBS). 16,23) Murine osteosarcoma LM8 cells 25) were grown and maintained at 37°C (5% CO 2 in air) in MEM-α (Wako Pure Chemical Industries, Ltd., Osaka, Japan) supplemented with 10% FBS. 26) Human T-lymphocyte Jurkat cells were maintained at 37°C (5% CO 2 ) in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% (v/v) FBS.…”

“…16) Statistical Analysis Welch's t-test (two-tailed) was performed for the statistical analyses. Values were considered to be significantly different when the p value was less than 0.05.…”

“…To date, we and others have shown that DIF-1, DIF-3, and their derivatives suppress cell growth and occasionally induce or promote cell differentiation in both cancer cells and non-transformed (normal) mammalian cells. [6][7][8][9][10][11][12][13][14][15] In addition, some DIF derivatives promote glucose consumption in mammalian cells in vitro and in vivo [16][17][18] ; others regulate interleukin 2 (IL-2) production in human Jurkat T cells [19][20][21] and suppress innate immune responses in Drosophila S2 cells. 21) Furthermore, some derivatives of DIF-3, specifically 1-(3-chloro-2,6-dihydroxy-4-methoxyphenyl)-heptan-1-one (DIF-3(+2)) and 1-(3-chloro-2,6-dihydroxy-4-ethoxyphenyl)hexan-1-one (Bu-DIF-3) (Fig.…”

A flavin-dependent halogenase catalyzes the chlorination step in the biosynthesis of Dictyostelium differentiation-inducing factor 1

“…DIF-1 biosynthesis and activity has been a long-standing topic of interest due to its essential role in the proper developmental program of Dictyostelium. DIF-1 has also garnered interest as a lead compound with various biological activities in mammalian cells (12)(13)(14).…”