engleski

Quantitative tandem mass tags proteomics of CT26.WT mouse colon cancer tissue after treatment with blended medicinal mushroom preparations with or without 5-fluorouracil

Abstract: Medicinal mushrooms are for millennia widely appreciated as natural sources of bioactive substances and are recognized as significant immunomodulatory and antitumor agents. In the last 50 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 organisms. 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) on quantitative changes in the tumor tissue proteome obtained by quantitative proteomic tandem mass tags approach using high-resolution Orbitrap technology. The 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. The study was conducted on Balb/c mice, 2-3 months old, average weight 20-25 grams. Seven groups of animals (control and 6 various combinations of medicinal mushroom preparations and 5-FU) were inoculated with 1 x 106 CT26.WT tumor cells and treated with the aforementioned substances for a period of 10 days afterwards. After isolation of protein from tumor tissue and determination of protein concentration in tissue lysates, enzymatic digestion of total isolated tissue proteins was performed. Subsequently, identification of separated proteins by means of tandem mass spectrometry (MS/MS) and search of specialized databases was performed. The analysis of differences in relative expression of proteins isolated from tissue samples was made by means of labeling with stable isotopes. Protein identification and relative quantification were performed using Proteome Discoverer, and data were analysed using R. Finally, the search of proteomic functional databases and literature references based on the results of mass spectrometry analysis was performed for the purpose of determining new potential protein candidates and biological processes which are affected by the treatment.

medicinal mushrooms ; polysaccharides ; CT26.WT ; Balb/c ; proteomics

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