engleski

The effects of blended medicinal mushroom preparations on survival and immunological and antimetastatic parameters in mouse CT26.WT colon cancer

INTRODUCTION Medicinal mushrooms are for millennia widely appreciated as natural sources of bioactive substances and are recognized as significant immunomodulatory and antitumor agents [1]. In the last 30 years there has been an increased interest in the use of fungi in the (traditional) medicine as well as in determining their therapeutic and medicinal properties, resulting in exponential growth of the number of scientific research papers on their impact on animal and human organism. This paper describes the effects of two commercial preparations from medicinal mushrooms, tableted preparation AGARIKON.1 and liquid preparation AGARIKON PLUS (manufacturer: Dr Myko San – Health from Mushrooms). These extract preparations from various mushroom species with proven antitumor effects were compared in various combinations, with or without 5-fluorouracil (5-FU), which is commonly used antimetabolite class drug used in treating colon cancer. An important hypothesis throughout is that the blend of different medicinal mushroom substances of the same class of compounds achieved synergistic effects, and consequently significantly stronger cytotoxic and other effects [2]. Moreover, since the preparations AGARIKON.1 and AGARIKON PLUS differ significantly in their polysaccharide : polyphenol contents ratio, this contents is then correlated with the observed Balb/c mice survival, as well as with other immunological and antimetastatic parameters [3, 4]. Besides observed effects on a) survival, we also record the effects of mushroom preparations with or without cytostatic drug on b) tumor growth, c) angiogenesis and tumor invasiveness, d) gene expression of chosen candidates and e) immunological parameters (macrophage function and cytokine expression). METHODS The study was conducted on Balb/c mice, 2-3 months old, average weight 20-25 grams. The first set of eight groups of animals (control and 7 various combinations of medicinal mushroom preparations and 5-FU) were inoculated with CT26.WT tumor cells [5] and, after a period of 2 weeks, treated with the aforementioned substances for a period of 2 weeks. The survival was observed for a period of 55 days from tumor inoculation, and results were presented using Kaplan-Meier estimator. The second set of eight groups of animals were treated as above, but sacrificed immediately after the end of the treatment period. MMP-2, MMP-9 ELISA tests were performed for the analysis of antimetastatic effects, Th1/Th2/Th17 cytokine panels (ELISA), NO (Griess reagent) and arginase for the analysis of the immunomodulatory effects. Angiogenesis and proliferation index was analyzed by immunohistochemistry (CD31 and Ki-67) and VEGF expression (ELISA). Anti-inflammatory effects were measured using COX expression (ELISA). Gene expression pertaining to macrophage activation and anti-inflammatory effects was confirmed by Western blot (NOS-2, COX-2) RESULTS AND DISCUSSION The results show significant beneficial effects on various parameters related to tumor suppression and show that medicinal mushrooms preparations alone and in combination with standard therapies have a major role in this in vivo model of colorectal cancer, which should be further explored in the clinic. References [1] Wasser SP (2011), Appl Microbiol Biotechnol 89, 1323-1332 [2] Jakopovich I (2011), Int J Med Mushr 13, 307-313 [3] Ehrke MJ (2003), Int Immunopharmacol 3, 1105-1119 [4] Durgo K, Koncar M, Komes D, Belscak-Cvitanovic A, Franekic J, Jakopovich I, Jakopovich N, Jakopovich B (2013), Int J Med Mushr 15, 435-48 [5] Castle J, Loewer M, Boegel S, Graaf de J, Bender C, Tadmor A, Boisguerin V, Bukur T, Sorn P, Paret C, Diken M, Kreiter S, Tureci O, Sahin U (2014), BMC Genomics 15, 190

medicinal mushrooms ; polysaccharides ; CT26.WT ; Balb/c ; immunomodulation ; metastasis

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