RRC ID |
52060
|
著者 |
Wakuri S, Yamakage K, Kazuki Y, Kazuki K, Oshimura M, Aburatani S, Yasunaga M, Nakajima Y.
|
タイトル |
Correlation between luminescence intensity and cytotoxicity in cell-based cytotoxicity assay using luciferase.
|
ジャーナル |
Anal Biochem
|
Abstract |
The luciferase reporter assay has become one of the conventional methods for cytotoxicity evaluation. Typically, the decrease of luminescence expressed by a constitutive promoter is used as an index of cytotoxicity. However, to our knowledge, there have been no reports of the correlation between cytotoxicity and luminescence intensity. In this study, to accurately verify the correlation between them, beetle luciferase was stably expressed in human hepatoma HepG2 cells harboring the multi-integrase mouse artificial chromosome vector. We showed that the cytotoxicity assay using luciferase does not depend on the stability of luciferase protein and the kind of constitutive promoter. Next, HepG2 cells in which green-emitting beetle luciferase was expressed under the control of CAG promoter were exposed to 58 compounds. The luminescence intensity and cytotoxicity curves of cells exposed to 48 compounds showed similar tendencies, whereas those of cells exposed to 10 compounds did not do so, although the curves gradually approached each other with increasing exposure time. Finally, we demonstrated that luciferase expressed under the control of a constitutive promoter can be utilized both as an internal control reporter for normalizing a test reporter and for monitoring cytotoxicity when two kinds of luciferases are simultaneously used in the cytotoxicity assay.
|
巻・号 |
522
|
ページ |
18-29
|
公開日 |
2017-4-1
|
DOI |
10.1016/j.ab.2017.01.015
|
PII |
S0003-2697(17)30037-4
|
PMID |
28111305
|
MeSH |
Animals
Chromosomes, Artificial, Mammalian* / genetics
Chromosomes, Artificial, Mammalian* / metabolism
Coleoptera
Hep G2 Cells
Humans
Insect Proteins* / genetics
Insect Proteins* / metabolism
Luciferases* / genetics
Luciferases* / metabolism
Luminescent Measurements / methods*
Mice
Promoter Regions, Genetic*
Toxicity Tests / methods
|
IF |
2.877
|
引用数 |
7
|
リソース情報 |
ヒト・動物細胞 |
Hep G2(RCB1886) |