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Expression and purification of recombinant human CCL5 and its biological characterization.
- Source :
-
The protein journal [Protein J] 2022 Apr; Vol. 41 (2), pp. 337-344. Date of Electronic Publication: 2022 May 07. - Publication Year :
- 2022
-
Abstract
- C-C motif chemokine ligand 5 (CCL5) is crucial in the tumor microenvironment. It has been previously reported to act as a key role in tumor invasion and metastasis. However, the function of exogenous CCL5 in ovarian cancer has not been well-characterized. The present study attempted to express and purify recombinant CCL5 protein and investigate the exogenous CCL5 in ovarian cancer cell proliferation. The human CCL5 was amplified and inserted into the pET-30a vectors for prokaryotic expression in Escherichia coli BL21. Soluble His-CCL5 was successfully expressed with 0.1 mmol/L of isopropyl-β-D-1-tiogalactopiranoside at 25 ℃ and purified by affinity chromatography. Additionally, methyl thiazolyl tetrazolium (MTT) assay demonstrated that CCL5 promotes ovarian cancer cell proliferation; increases the phosphorylation levels of extracellular-signal-regulated kinase and mitogen-activated protein kinase/ERK kinase, and increases the mRNA levels of Jun, NF-κB2, Nras, Relb, and Traf2. Furthermore, treatment with the MEK inhibitor reduced the Jun, NF-κB2, and Traf2 mRNA levels, indicating that exogenous CCL5 increased ovarian cancer cell proliferation, through MEK/ERK pathway activation, and Jun, NF-κB2, and Traf2 expression. The present study provided primary data for further studies to discover more CCL5 functions in ovarian cancer.<br /> (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
- Subjects :
- Cell Line, Tumor
Chemokine CCL5 genetics
Chemokine CCL5 metabolism
Chemokine CCL5 pharmacology
Female
Humans
Mitogen-Activated Protein Kinase Kinases metabolism
RNA, Messenger metabolism
TNF Receptor-Associated Factor 2 metabolism
Tumor Microenvironment
NF-kappa B p52 Subunit
Ovarian Neoplasms genetics
Ovarian Neoplasms metabolism
Ovarian Neoplasms pathology
Subjects
Details
- Language :
- English
- ISSN :
- 1875-8355
- Volume :
- 41
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- The protein journal
- Publication Type :
- Academic Journal
- Accession number :
- 35524873
- Full Text :
- https://doi.org/10.1007/s10930-022-10047-8