Antiproliferative activity in vitro and in vivo of the spicamycin analogue KRN5500 with altered glycoprotein expression in vitro.
The spicamycin analogue KRN5500 (NSC 650426; SPA) is derived from Streptomyces alanosinicus. The unique structure contains a purine, an aminoheptose sugar, glycine, and a tetradecadiene fatty acid. SPA potently inhibits the growth of certain human tumor cell lines in vitro (IC50 for growth <100 nM) and displays marked activity in vivo in Colo 205 colon carcinoma xenografts. Selective inhibition of labeled precursor incorporation was not evident at 1 or 4 h of exposure to the drug, but at 8 h, [3H] leucine incorporation was inhibited by approximately 40% at or below the IC50 for cell growth. Because of the structural similarity of SPA to inhibitors of glycoprotein processing, we examined the effect of SPA on indicators of glycoprotein synthesis and processing in HL60TB promyelocytic leukemia and Colo 205 colon carcinoma cells. Brief periods of exposure ( approximately 30 min) to SPA at the IC50 for growth increased incorporation of [3H]mannose. When examined by Western blotting after prolonged (40-48 h) incubation with lectins that target mannose-containing carbohydrates, Galanthus nivalis agglutinin and concanavalin A, a qualitative change in the pattern of mannose-containing glycoproteins was observed in HL60TB cells. Significant changes in the pattern of surface glycoprotein expression in intact cells were demonstrated by flow cytometry using fluorescence-labeled lectins. An increase in the number of cells binding G. nivalis agglutinin (indicating terminal mannose) was noted, but a decrease in the amount of lectin bound per cell was noted for wheat germ agglutinin (detecting sialic acid and terminal beta-N-acetyl glucosamine residues). Electron microscopy revealed loss of microvilli, and the Golgi apparatus appeared inflated. Our findings, therefore, raise the possibility that cells exposed to SPA have altered glycoprotein processing after exposure to low concentrations of drug, prior to the occurrence of overt cytotoxicity. These effects are consistent with a prominent early effect of SPA on the enzymatic machinery or organelles important for proper glycoprotein processing and emphasize the novelty of this agent's likely mechanism of action.
Burger
A M
AM
Biological Testing Branch, Laboratory of Pharmaceutical Chemistry, Frederick Cancer Research and Development Center, Frederick, MD 21702-1201, USA.
Kaur
G
G
Hollingshead
M
M
Fischer
R T
RT
Nagashima
K
K
Malspeis
L
L
Duncan
K L
KL
Sausville
E A
EA
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000627:therapeutic use / Q000633:toxicity*; D002455:Cell Division / Q000187:drug effects*; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000473:pathology / Q000648:ultrastructure; D004354:Drug Screening Assays, Antitumor; D005434:Flow Cytometry; D027801:Galanthus; D006023:Glycoproteins / Q000096:biosynthesis*; D018922:HL-60 Cells; D006801:Humans; D037102:Lectins; D008358:Mannose / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D051379:Mice; D008819:Mice, Nude; D037121:Plant Lectins; D011684:Purine Nucleosides / Q000627:therapeutic use / Q000633:toxicity; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D000903:Antibiotics, Antineoplastic; D006023:Glycoproteins; D037102:Lectins; D008562:Membrane Glycoproteins; D037121:Plant Lectins; D011684:Purine Nucleosides; C092296:KRN 5500; D008358:Mannose