The Antiproliferative, Proapoptotic and Antitumour Activity in Vitro of 10% PBS Holocene Grain Washout Against CaCo-2 CELLS can be Enhanced with Different Human Alfa Interferons

Holocene sands near river Drava (Croatia) contain Holocene minerals which, in grained form, exhibit unusual biological/microbiological activity. The grained Holocene minerals were analysed : SiO₂ (88,71%), Al₂O₃(5,37%), Fe₂O₃ (1,08%) and gold (Au) (12,1% ) among elements. Other elements were present in smaller range. It was found that a 10% Holocene grain Phosphate Buffer Saline (PBS) washout and HuIFN – αN3 affect the growth of CaCo-2 cells . The aim was to measure the effect of different Human Alfa IFNs on antiproliferative, proapoptotic and antitumour activity in vitro of 10% PBS washout of Holocene grain on CaCo-2 cells . The following interferons were used: HuIFN-αN3 (IMZ – Zagreb, Croatia), Alfaferon (Alfa Wasserman, Italy), Egiferon (Trigon, Hungary); Human recombinant αA Interferon (EMD Chemicals Inc., San Diego,Ca 92121 USA), Roferon-A (IFN Alfa-2a), Roche and Intron A (IFN Alfa-2b) (MSD Italia Srl, Italy). The interferons were analysed by RP-HPLC at 280 nm. CaCo-2 cells were treated as: (a) cell control, (b) interferon + 10% PBS (1:1,1:2,2:1), (c) interferon, (d) 10% PBS. Comparison with the interferon-untreated cells was calculated. The apoptotic cells were isolated by BioVision: Apoptotic cell isolation kit. The 10% Holocene grain washout showed an antiproliferative activity. This can be enhanced up to three times by HuIFN-αN3 and not with the HuIFN-α2a or HuIFN-α2b. In case of proapoptotic activity, the 10% Holocene grain washout shows 26,52% of apoptotic cells. It was increased to 49,85% with HuIFN-αN3. With HuIFN-α2a and HuIFN-α2b it was 22,80% and 42,60%, respectively. It can be concluded that 10% PBS Holocene grain washout synergizes the effect of the HuIFN-αN3 in highest level of AP activity, while this is not the case with HuIFN-α2a or HuIFN-α2b.