Purification and Characterization of a New D-Galactose-Specific Lectin from the Housefly,Musca domestica, and Its Antiproliferative Effect on Human K562 and MCF-7 Tumor Cells

Abstract

In the present work, a D-galactose-specific lectin with novel N-terminal sequence was purified from Musca domestica L. (Diptera: Muscidae) pupae. The purification was performed using affinity chromatography, ultra-filtration, and HPLC. The haemagglutinating activity of M. domestica lectin was specifically inhibited by D-galactose. The haemagglutinating activity of this lectin was stable at temperatures up to 65° C and in pH ranging from 4 to 8. Salts including FeCl3 and MnCl2 inhibited the haemagglutinating process, whereas NaCl, KCl, CaCl2, MgCl2, ZnCl2, and AlCl3 did not. By SDS-PAGE, purified M. domestica pupae lectin yielded a single band with a molecular weight of 40 kDa, with or without reduction of β-mercaptoethanol, and it could be stained with Alcian Blue 8 GX. The morphology of purified lectin was observed by atomic force microscopy, which indicated that M. domestica lectin was an 8.27 nm high, globular shaped glycoprotein with a 1.41 nm high polysaccharide chain. In addition, antiproliferative activity of this lectin against tumor cells K562 and MCF-7 was determined with a colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, which showed that the antiproliferative process was time- and dose-dependent with an IC50 of 5.7 and 6.7 at 24 h, 5.5 and 6.4 at 36 h, 5.2 and 6.5 µM at 48 h, respectively.

Musca domestica Pupae Lectin Improves the Immunomodulatory Activity of Macrophages by Activating Nuclear Factor-κB

“…Lectins isolated from various Lepidoptera insects have been shown to profoundly affect the immune system in vivo and in vitro. [28][29][30][31] In the present study, immunomodulatory effects of MPL isolated from M. domestica pupae, which inhibited the viability of human breast cancer (MCF-7) cells and human leucocythemia (K562) cells, 24 were investigated in vitro. It is well known that as phagocytic cells, macrophages are widely distributed throughout the body and play important roles in host defense by coordinating innate immunity and …”

“…[21][22][23][24] Although it is well known that macrophages play a key role in host defense mechanisms, the immunoregulatory function of purified MPL on macrophages remains to be reported. The aim of the present study was to investigate the immunomodulatory effects of MPL extracted from M. domestica pupae on murine peritoneal macrophages in vitro.…”

Lectin purified from Musca domestica pupa up-regulates NO and iNOS production via TLR4/NF-κB signaling pathway in macrophages

“…In our laboratory, lectin from M. domestica pupa had also been shown to upregulate the production of NO without clarifying the mechanism behind the scenes. In our previous study, the MPL was purified from M. domestica pupa and it had been demonstrated that MPL could inhibit the viability of human breast cancer (MCF-7) cells and human leucocythemia (K562) cells [19]. In this study, the MPL was examined by MTT assay and Griess reaction, which showed that MPL increased NO production in macrophage without exhibiting cell toxicity.…”

“…They can also increase immunomodulatory activity in macrophages by enhancing the phagocytosis and the secretion of TNF-α and NO [16][17][18]. In our preliminary studies, one novel lectin purified from M. domestica pupa was identified with its biochemical properties, morphology and N-terminal sequence [19] and it has been demonstrated that the other lectin also purified from M. domestica pupa has the immunomodulatory activity in macrophages and increases the production of NO [20]. However, the mechanism that lectins induce NO production in macrophages is not reported.…”

A lectin from Musca domestica pupae stimulates B cell proliferation and enhances IL-12 production via ERK1/2-NF-κB signaling pathways

“…In our previous study, some lectins have been isolated and characterized from Musca domestica pupae. The lectin of 40 kDa is an 8.27 nm, globular glycoprotein with a 1.41 nm polysaccharide chain, which can inhibit proliferative activity of tumor cells K562 and MCF-7 (Cao et al 2008(Cao et al , 2009). However, the function of this lectin on B cells and the mechanism is still unclear.…”