Open AccessA simple in vitro tumor chemosensitivity assay based on cell penetrating peptide tagged luciferase
Author(s) -
Tingyu Yu,
Jiao Lin,
Jin Zhao,
Wei Huang,
Lingwen Zeng,
Zhiyuan Fang,
Ning Xu
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0186184
Subject(s) - luciferase , intracellular , chemosensitivity assay , extracellular , cell culture , microbiology and biotechnology , cell , lysis , recombinant dna , cancer cell , cytotoxicity , in vitro , biology , transfection , chemistry , cancer research , biochemistry , cancer , gene , genetics
The analysis of intracellular ATP can reveal the response of cells to different treatments and is important for individualized medicine. In the present study, we developed a cell penetrating peptides (CPPs) tagged luciferase (TAT-LUC) for tumor chemosensitivity assay. The activity of recombinant TAT-LUC was evaluated using ATP standard solution and tumor cells. This recombinant TAT-LUC was then used for the analysis of sensitivity index (SI) of four strains of tumor cells. The results showed that TAT-LUC could detect less than 10 nM extracellular ATP with a strong correlation between the luminescence intensity and the ATP content (R 2 = 0.994). Without cell lysis, the detection limit for intracellular ATP analysis was 40 tumor cells. Furthermore, chemosensitivity of four strains of tumor cells (Skov-3/DDP, A549/DDP, MDA-MB-231, Huh-7) was determined by this assay successfully. The cell penetration ability of TAT-LUC enables the assay not only to reflect drug resistance of tumor cells real-timely but also to minimize the test time, which can be a valuable aid for personalized cancer chemotherapy.