The purpose was to investigate the influence of RJ-F(M), 10-hydroxy-2decenoic acid and HuIFN-αN3 on the proliferation of CaCo-2 cells and ascertain their effects on intracellular glutathione level and lipid peroxidation. The antiproliferative (AP) activity of RJ-F (M) (0.1 g/10 mL PBS), HuIFN-αN3 (1000 IU mL −1), 10-HDA (100.0 μmol L −1) and their combinations, in the ratios 1:1, 1:2, and 2:1 on CaCo-2 cells were measured. Single RJ-F (M) had a low AP activity: 2.0 (0.5 mg mL −1). HuIFN-αN3 had an AP activity of 2.5 (208.33 IU mL −1), while 10-HDA had an AP activity of 1.5 (37.5 μmol mL −1). AP activity of 3.8 was obtained when RJ-F(M) and HuIFN-αN3 were in the ratio 2:1. In it, the level of GSH was 24.9 ± 2.4 nmol g −3 of proteins (vs. 70.2 ± 3.2 nmol g −3 in the control), and level of MDA was 72.3 ± 3.1 nmol g −3 (vs. 23.6 ± 9.1 nmol g −3 in the control). 10-HDA, together with HuIFN-αN3, is responsible for the inhibition of CaCo-2 cell proliferation in vitro. RJ-F (M) and HuIFN-αN3 applied at 2:1 decreased level of GSH and increased lipid peroxidation via MDA in CaCo-2 cells. Future studies are needed whether these GSH-and MDA-related activities of RJ-F (M), HuIFN-αN3, 10-HDA, and their combinations may decrease the tumorigenicity index and tumorigenic potential of tumor cells in vitro

Among royal jelly's (RJ) various biological activities, its possible antitumour activity deserves particular attention. The purpose of this study was to investigate the influence of RJ, its bioactive component 10-hydroxy-2-decenoic acid (10-HDA), and human interferon-alpha (HuIFN-αN3) on the proliferation of human colorectal adenocarcinoma cells (CaCo-2), and ascertain their effect on intracellular glutathione (GSH) level and lipid peroxidation. We studied the antiproliferative (AP) activity of RJ [(0.1 g/10 mL phosphate buffer saline (PBS)], HuIFN-αN3 (1000 I.U. mL , and their different combinations, in the ratio 1:1, 1:2, and 2:1 on CaCo-2 cells. The GSH level was measured by glutathione assay. The lipid peroxidation was measured by malondialdehyde (MDA) assay. Single RJ had a low AP activity: 2.0 (0.5 mg mL -1 ). HuIFN-αN3 had an AP activity of 2.5 (208.33 I.U. mL -1 ), while 10-HDA had an AP activity of 1.5 (37.5 μmol mL). The highest AP activity of 3.8 was obtained when RJ and HuIFN-αN3 were applied at the ratio 2:1. In that combination the level of GSH was 24.9±2.4 nmol g -3 of proteins (vs. 70.2±3.2 nmol g -3 in the control) and the level of MDA was 72.3±3.1 nmol g -3 (vs. 23.6±9.1 nmol g -3 in the control). It is generally assumed that 10-HDA, an important constituent of RJ, together with HuIFN-αN3, is responsible for the inhibition of CaCo-2 cells proliferation in vitro. In our study, however, RJ and HuIFN-αN3 applied at 2:1 decreased the level of GSH the most and significantly increased lipid peroxidation via MDA in CaCo-2 cells. Future studies should show whether these GSH-and MDA-related activities of RJ, HuIFN-αN3, 10-HDA, and their combinations may decrease the tumorigenicity index and tumorigenic potential of various tumour cells in vitro. KEY WORDS: antiproliferative activity; antitumour activity; CaCo-2 cells; 10-hydroxy-2-decenoic acid; malondialdehydeRoyal jelly (RJ) is a milky material secreted by the hypopharyngeal and mandibular glands of young worker bees between the sixth and twelfth day of their life. It is food exclusively for the queen honey bee (Apis mellifera) larva, which leads to the development of a sexually mature queen bee (1, 2). Chemically, RJ comprises water (50-60 %), different proteins (18.0 %), carbohydrates (15.0 %), lipids (3-6 %), mineral salts (1.5 %), and vitamins, together with a large number of bioactive substances, such as 10-hydroxy-2-decenoic (10-HDA) acid (3). RJ exhibits various immunomodulatory and antiproliferative properties, as well as possible antitumour activity. It also contains different antibacterial proteins like 350-kDa protein stimulating the proliferation of human monocytes (4-6). In addition, the RJ Protein 30 fraction exhibits a clear cytotoxic effect on HeLa cells by decreasing the initial cell population by 50 % at the end of treatment (7).Impact of royal jelly and HuIFN-αN3 on proliferation, glutathione, and lipid peroxidation in human colorectal adenocarcinoma cells in vitro Arh Hig Rada Toksikol 2015;66:269-274

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