Antitumour activity of Helix hemocyanin against bladder carcinoma permanent cell lines

Abstract

Hemocyanins are oxygen-transporting glycoproteins in molluscs and arthropods. In this study, we assayed the biomacromolecules from the molluscs Helix lucorum (HlH), Rapana venosa (RvH) and Megatura crenulata (KLH) including their functional units (FUs), for therapy of bladder cancer permanent cells. In vitro studies antitumour activities of these proteins were performed with bladder cancer permanent cell line CAL-29 and the normal urothelial cell line HL 10/29 in comparison to doxorubicin. The obtained results showed that the human tumour CAL-29 cell line is sensitive to the action of the tested hemocyanins and their isoforms. We observed a dose-and time-dependent inhibition of tumour cell growth after incubation with native HlH and two FUs (b c-HlH-a and FU b c-HlH-h); and of particular significance, FU b c-HlH-h showed a surprisingly stronger effect than that the doxorubicin-treated cells. Cells treated with b c-HlH-h, showed both, apoptotic and necrotic cells. In addition, two-dimensional polyacrylamide gel electrophoresis (PAGE) found for differential up-regulation of several proteins after hemocyanin treatment. Eight different down-regulated and two up-regulated proteins were identified, which may be associated with the apoptosis pathway. No inhibition of the normal urothelial cell line HL 10/29 was observed after treatment with HlH and its isoforms. The most effective inhibition of CAL-29 tumour cells was observed after treatment with b c-HlH-h, probably caused by a specific oligosaccharide structure of HlH with methylated hexoses. These results suggest that hemocyanin glycosylation plays an important role in its anticancer activity.

Hemocyanins from Helix and Rapana Snails Exhibit in Vitro Antitumor Effects in Human Colorectal Adenocarcinoma

“…The search for novel, more effective, and safe antitumor medicines with natural origin is one of the main trends of contemporary oncology research. In this respect, hemocyanin oligomeric copper-containing glycoproteins that function as oxygen carriers in the hemolymph of mollusk and arthropod species represent significant interest because they combine strong immunostimulating activity and direct anticancer effects [ 5 , 6 , 25 , 26 , 27 ]. Among this class of compounds, the hemocyanin obtained from Megathura crenulata , known as keyhole limpet hemocyanin (KLH), has been most extensively studied and has found a number of biotechnological and medical applications.…”

Hemolymph of molluscan origin: from biochemistry to modern biomaterials science

“…Recently, molluscan hemocyanin has found applications as an antibacterial and antiviral factor owing to its potential phenoloxidase activity. These effects have been demonstrated in a number of reports [30,31,51,75,101]. H. pomatia hemocyanin was successfully used as a novel bioadjuvant for viral and bacterial antigens [101].…”

“…Such terrestrial gastropod species as Achatina fulica (Bowdich, 1822), Cornu aspersum (ex. Helix aspersa) (Müller, 1774) and the aquatic mollusk species as Rapana venosa (Valenciennes, 1846), Megathura crenulata (Sowerby, 1825) are only a few selected examples from which hemolymph is used as a source of such biologically active substances [25][26][27][28][29][30][31][32][33].…”

An 1H NMR- and MS-Based Study of Metabolites Profiling of Garden Snail Helix aspersa Mucus

“…In contrast to other hemocyanins, three isoforms (β-, αN- αD-HaH, and β-, αN- αD-HlH) with a molecular weight (Mw) of about 450 kDa were isolated from both hemocyanins Helix lucorum and Helix aspersa , respectively [ 7 ]. Hemocyanins rom Helix lucorum and Helix aspersa can be carriers of poorly immunogenic antigens, and are promising adjuvants for viral and bacterial vaccines and their use as novel antitumor, antibacterial, and immunotherapeutic agents has also been established [ 13 , 14 , 15 ].…”