[Cloning and expression of human interleukin 24 gene and study on its activity of inducing tumor cells apoptosis in vitro].
OBJECTIVE
To clone and express IL-24 gene in E.coli and to study its antitumor activity to induce apoptosis of tumor cells after purification and refolding.


METHODS
The total RNA was extracted from peripheral blood mononuclear cells (PBMC) and induced by ConA. The human IL-24 gene was amplified by RT-PCR before subcloned into expression vector pET32a(+) and expressed in E. coli. The fusion protein was identified by Edman and purified in chelating sepharose fast flow chromatography and refolding. The activity of inducing tumor cells apoptosis was observed by MTT colorimetry and morphological study in vitro.


RESULTS
The obtained human IL-24 gene was identical with that included in GenBank. The recombinant IL-24 could be expressed in E. coli by IPTG induction as inclusion body (35 kD) with purity over 95%. The apoptosis of MCF-7 cells was induced by Trx-IL-24 protein.


CONCLUSIONS
The fusion protein can be highly expressed in E. coli and the obtained protein with high purity can induce tumor cells apoptosis.
Yang
Jun
J
Key Laboratory of Biomechanics and Tissue Engineering, Chongqing University, Chongqing 400044, China. w8301991@263.net
Cai
Shao-xi
SX
Zou
Quan-ming
QM
Zhang
Wei-jun
WJ
Yang
Qin
Q
D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods*; D006801:Humans; D007378:Interleukins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099180:Interleukin-24
D007378:Interleukins; D000099180:Interleukin-24