The Influence of Spirulina platensis Filtrates on Caco-2 Proliferative Activity and Expression of Apoptosis-Related microRNAs and mRNA

Abstract

Spirulina platensis (SP) is a blue-green microalga that has recently raised attention not only as a nutritional component, but also as a source of bioactivities that have therapeutic effects and may find application in medicine, including cancer treatment. In the present study we determined the cytotoxic effect of S. platensis filtrates (SPF) on human colon cancer cell line Caco-2. Three concentrations of SPF were tested—1.25%, 2.5%, and 5% (v/v). We have found that the highest concentration of SPF exerts the strongest anti-proliferative and pro-apoptotic effect on Caco-2 cultures. The SPF negatively affected the morphology of Caco-2 causing colony shrinking and significant inhibition of metabolic and proliferative activity of cells. The wound-healing assay showed that the SPF impaired migratory capabilities of Caco-2. This observation was consistent with lowered mRNA levels for metalloproteinases. Furthermore, SPF decreased the transcript level of pro-survival genes (cyclin D1, surviving, and c-Myc) and reduced the autocrine secretion of Wnt-10b. The cytotoxic effect of SPF involved the modulation of the Bax and Bcl-2 ratio and a decrease of mitochondrial activity, and was related with increased levels of intracellular reactive oxygen species (ROS) and nitric oxide (NO). Moreover, the SPF also caused an increased number of cells in the apoptotic sub-G0 phase and up-regulated expression of mir-145, simultaneously decreasing expression of mir-17 and 146. Obtained results indicate that SPF can be considered as an agent with anti-cancer properties that may be used for colon cancer prevention and treatment.

Nanocrystalline Hydroxyapatite Loaded with Resveratrol in Colloidal Suspension Improves Viability, Metabolic Activity and Mitochondrial Potential in Human Adipose-Derived Mesenchymal Stromal Stem Cells (hASCs)

“…For the analysis of mRNA expression, cDNA was obtained from 1 µg of RNA using transcription Tetro cDNA Synthesis Kit (Bioline Reagents Ltd., London, UK). The synthesis of cDNA used for microRNA detection was performed using Mir-X miRNA First-Strand Synthesis Kit (Takara Clontech, Biokom, Janki, Poland) according to the protocol described previously [44,45]. The reactions of reverse transcription were performed on T100 Thermal Cycler (Bio-Rad Polska Sp.…”

<p>Zirconium Oxide Thin Films Obtained by Atomic Layer Deposition Technology Abolish the Anti-Osteogenic Effect Resulting from miR-21 Inhibition in the Pre-Osteoblastic MC3T3 Cell Line</p>

“…The morphology of cells was investigated after 96 h using well-described protocols. 31,32 In order to visualize the mitochondria, cells were stained with MitoRed dye (Sigma Aldrich Sp. z o.o., Poznan, Poland) prepared in CGM at concentration (1:1000).…”

“…The details of the protocols were described earlier. 35,36 The reverse transcription was performed using a Tetro cDNA Synthesis Kit (Bioline Reagents Ltd., London, United Kingdom) according to the protocol provided by the manufacturer. Additionally, cDNA was synthesized from 375 ng of total RNA using Mir-X™ miRNA First-Strand Synthesis Kit (Takara Bio Europe, Saint-Germain-en-Laye, France).…”

Spirulina extract enriched for Braun‐type lipoprotein (Immulina®) for inhibition of 4T1 breast tumors' growth and metastasis

“…These results indicate Spirulina extract can directly induce apoptosis to the cancer cells which is in consistence with previous studies. (Czerwonka et al, ; Śmieszek et al, )…”