RESUMO
BACKGROUND: 3D cancer stem cell (CSC) cultures are widely used as in vitro tumor models. In this study, we determined the effects of enriching HCT116 tumor spheres initially cultured in serum-free medium with different concentrations of serum, focusing on the effect of microserum environment stimulation on extraction and biological function of colorectal cancer stem cells (CCSCs). METHODS: CCSCs were enriched in standard serum-free medium and serum-free medium with different concentrations of serum for 1 week. The expression of CSC-associated markers in CCSCs, and the presence and relative proportion of CSCs (CD133/CD44 cell sorting) were then determined to elucidate the effect of the microserum environment on the preservation of CSC-related features. Further, the tumorigenic capacity of CCSCs was evaluated in an immunodeficiency mouse model. RESULTS: Our data indicated that a significantly greater number of spheres with a greater size range and high viability without drastic alteration in biological and structural features, which maintained self-renewal potential after sequential passages were formed after serum supplementation. Real-time analysis showed that both serum spheres and serum-free spheres displayed similar expression patterns for key stemness genes. Serum spheres showed higher expression of the CSC surface markers CD133 and CD44 than did CSCs spheres cultured in serum-free medium. Adherent cultures in complete medium could adapt to the serum-containing microenvironment faster and showed higher proliferation ability. The addition of serum induced EMT and promoted the migration and invasion of serum globular cells. Compared with serum-free cells and adherent cells, serum spheres showed higher tumor initiation ability. CONCLUSIONS: Microserum environment stimulation could be an effective strategy for reliable enrichment of intact CCSCs, and a more efficient CSC enrichment method.
RESUMO
Background: Metagentiana rhodantha (Franch.) T.N.Ho and S.W.Liu (MR) belongs to Gentianales, and it is often called Hong-hua-long-dan in Chinese. Traditionally, it has been used to cure acute icteric hepatitis, sore throat, dysentery, acute gastritis, carbuncle, and furuncle based on traditional Chinese medicine (TCM) concepts. Aim of Study: This review manages to provide a critical and comprehensive analysis on the traditional uses, phytochemistry, pharmacology, toxicology, and clinical uses of MR and to evaluate the therapeutic potential of this plant. Methods: Relevant data mainly literatures on MR were selected from available database. All the papers reviewed provided evidence that the source herbs were reliably identified. Results: The heat-clearing and removing the phlegm, and purging fire and removing toxicity of MR contribute to its dispelling jaundice, and clearing lung heat and cough. The compounds isolated from this plant include iridoids and secoiridoids, phenolic acids, ketones, triterpenoids, flavonoids, benzophenone glycosides, and others. Mangiferin (MAF) is a characteristic substance from this plant. The pharmacological studies show that some extracts and compounds from MR exhibit anti-inflammatory, antinociceptive, antibacterial, hepatoprotective, cardioprotective, and other effects which are associated with the traditional uses of this plant. The toxicological studies suggest that MAF is less toxic in mice and dogs. Nowadays, Chinese patent drugs such as Feilike Jiaonang and Kangfuling Jiaonang containing MR have been used to cure cough, asthma, chronic bronchitis, dysmenorrhea, and appendagitis. Conclusion: Although the current studies provide related research information of MR, it is still necessary to systemically evaluate the chemistry, pharmacology, toxicity, and safety of the extracts or compounds from this plant before clinical trials in the future. In addition, except for lung infection-related diseases, analgesia, anti-tumor, and hypertriglycemia may be new and prior therapeutic scopes of this ethnomedicine in the future.