Quantitative Proteomic Profiling of Tachyplesin I Targets in U251 Gliomaspheres

Abstract

Tachyplesin I is a cationic peptide isolated from hemocytes of the horseshoe crab and its anti-tumor activity has been demonstrated in several tumor cells. However, there is limited information providing the global effects and mechanisms of tachyplesin I on glioblastoma multiforme (GBM). Here, by using two complementary proteomic strategies (2D-DIGE and dimethyl isotope labeling-based shotgun proteomics), we explored the effect of tachyplesin I on the proteome of gliomaspheres, a three-dimensional growth model formed by a GBM cell line U251. In total, the expression levels of 192 proteins were found to be significantly altered by tachyplesin I treatment. Gene ontology (GO) analysis revealed that many of them were cytoskeleton proteins and lysosomal acid hydrolases, and the mostly altered biological process was related to cellular metabolism, especially glycolysis. Moreover, we built protein–protein interaction network of these proteins and suggested the important role of DNA topoisomerase 2-alpha (TOP2A) in the signal-transduction cascade of tachyplesin I. In conclusion, we propose that tachyplesin I might down-regulate cathepsins in lysosomes and up-regulate TOP2A to inhibit migration and promote apoptosis in glioma, thus contribute to its anti-tumor function. Our results suggest tachyplesin I is a potential candidate for treatment of glioma.

Tachyplesin Causes Membrane Instability That Kills Multidrug-Resistant Bacteria by Inhibiting the 3-Ketoacyl Carrier Protein Reductase FabG

“…At a final concentration of 100 mg/L, tachyplesin caused lysis of approximately 2% of hemocytes ( Figure 2A ), as well as a decline in the cell viability of 293FT human embryonic kidney cells ( Figure 2C ) and a sharp decline in the cell viability of L919 murine fibroblasts ( Figure 2B ), a cell line commonly used for toxicity testing. Interestingly, tumor cells such as A549 adenocarcinomic human alveolar basal epithelial cells seem to be more resistant to tachyplesin ( Figure 2D ), which made the reports of its anticancer effects somewhat controversial ( Evans et al, 2017 ; Li et al, 2017 ).…”

“…Proteomic profiling showed that low concentrations of tachyplesin may promote apoptosis in the glioblastoma multiforme cell line U251 by affecting intracellular enzymes and cause cell death in E. coli by inactivating intracellular esterases ( Hong et al, 2015 ; Li et al, 2017 ). However, treatment with a high concentration of tachyplesin caused the rapid death of K562 human erythroleukemia cells and E. coli , accompanied by complete cell membrane rupture in both cases ( Paredes-Gamero et al, 2012 ; Hong et al, 2015 ).…”

“…Research using confocal microscopy and flow cytometry to investigate the anti-tumor activities of tachyplesin showed that it induced controlled cell death by promoting apoptosis of the human erythroleukemia cell line K562 at lower concentrations and resulted in cell membrane disruption at higher concentrations ( Paredes-Gamero et al, 2012 ). Proteomic profiling of the effects of tachyplesin on glioblastoma multiforme cell line U251 also implied that tachyplesin may cause cell death by affecting intracellular enzymes and by promoting apoptosis ( Li et al, 2017 ). Regarding its antimicrobial effects, confocal microscopy and flow cytometry showed that bacteria died gradually in response to a sublethal concentration of tachyplesin and died instantaneously upon exposure to high concentrations of tachyplesin.…”

Research Development on Horseshoe Crab: A 30-Year Bibliometric Analysis

“…In view of the mechanism of tachyplesin, more researchers should focus on the discrimination between bacterial and mammalian FabG proteins to select specific inhibitory ligands and develop safe antibiotics (Liu et al, 2018). Tachyplesin I from horseshoe crabs shows an antitumor activity; however, the action mode of tachyplesin I in tumor cells remains unclear (Li et al, 2017). The new antimicrobial peptide polyphemusin III can induce cell death by destroying the plasma membrane (Marggraf et al, 2018).…”

<p>Transcriptome Analysis Reveals the Resistance Mechanism of <em>Pseudomonas aeruginosa</em> to Tachyplesin I</p>

“…10 With potent and broad-spectrum activities against both Gram-positive and Gram-negative bacteria, tachyplesin I has been found as a promising candidate for the development of antiinfection, anti-tumor, and anti-virus drugs. [11][12][13] Bacterial resistance to tachyplesin I has been induced in Aeromonas hydrophila XS91-4-1, P. aeruginosa CGMCC1.2620 (PA1.2620), P. aeruginosa ATCC27853 (PA27853), Escherichia coli ATCC25922 and F41 with crossresistance to other AMPs (e.g., pexiganan, tachyplesin III, and polyphemusin I), conventional antimicrobial agents (amikacin), as well as the potential involvement of extracellular proteases in mediating tachyplesin I-resistance in Gram-negative bacteria reported in previous studies. 14,15 A limited number of researches concentrated on the mechanism of bacterial resistance to tachyplesin I.…”