Transport of trans-activator of transcription (TAT) peptide in tumour tissue model: evaluation of factors affecting the transport of TAT evidenced by flow cytometry

Abstract

Objectives: Trans-activator of transcription (TAT), a cell penetrating peptide, has been explored to overcome resistance to penetration and transport inside the cell, therefore, suggested to be used as drug delivery vector into drug-resistant tumours. The generosity of this study was to evaluate modifiable factors (concentration, temperature, incubation time and spheroid age) on the penetration of TAT. Methods: Multicellular tumour spheroids (MCTS) used as tumour tissue models to mimic some characteristics with in-vivo tumors. Cell monolayer and 3-, 5-, 7-day-old MCTS were incubated with TAT and effects of modifiable factors were determined quantitatively through flow cytometry, based on TAT-positive cell count (%) and mean fluorescence intensity. Key findings: Enhancing TAT concentration (1, 5 and 25 µm), transport significantly increased (ANOVA, P < 0.0001) in cell monolayer and spheroids. However, rising temperature from 7 to 37°C (t, P > 0.05) and increasing incubation time; 20 min, 1 h and 3 h; (ANOVA, P > 0.05) were statistically non-significant. Moreover, TAT penetration declines as spheroids get older (ANOVA, P < 0.01). Conclusion: While exploiting MCTS as tumour tissue model, older spheroids could be preferred to target penetration-resistant cells and mimic the in-vivo microenvironment.