Back to Search
Start Over
Downregulation of mouse CCR3 by lentiviral shRNA inhibits proliferation and induces apoptosis of mouse eosinophils
- Source :
- Molecular Medicine Reports
- Publication Year :
- 2016
- Publisher :
- D.A. Spandidos, 2016.
-
Abstract
- RNA interference has been considered as an effective gene silencing method in basic and preclinical investigations. The aims of the present study were to construct a lentiviral vector expressing a short hairpin RNA (shRNA) targeting the murine CC chemokine receptor 3 (mCCR3), and to investigate its effects on the proliferation and apoptosis of mouse eosinophils. A recombinant lentiviral vector expressing four fragments of mouse CCR3 shRNA (pLVX‑mCCR3‑1+2+3+4‑shRNA) was constructed using subcloning techniques. This novel lentivirus was then packaged into 293T cells by co‑transduction with plasmids, including Baculo p35, pCMV R8.2 and VSV. The interference effects of the vector were verified using polymerase chain reaction (PCR) and western blot analyses. The effects of the interference on the proliferation and apoptosis of mouse eosinophils were investigated using 3‑(4,5‑dimethylthiazol‑2‑yl)‑5‑(3‑carboxymethoxyphenyl)‑2‑(4‑sulfophenyl)‑2H‑tetrazolium and terminal deoxynucleotidyl transferase dUTP nick end labeling methods, respectively. The results of the PCR and western blot analyses confirmed that the novel recombinant vector, pLVX‑mCCR3‑1+2+3+4‑shRNA, had high efficiency in inhibiting the mRNA and protein expression levels of mCCR3 in mouse eosinophils. The downregulation of mCCR3 significantly inhibited proliferation of the eosinophils. Furthermore, the present study found that the downregulation of mCCR3 significantly promoted apoptosis of the eosinophils. Therefore, the downregulation of mCCR3 led to the inhibition of proliferation and induction of apoptosis in mouse eosinophils. The predominant characteristics of allergic rhinitis are eosinophil infiltration and release of inflammatory mediators, which appear in a variety of clinical manifestations. The results of the present study indicate that mCCR3 silencing may serve as a putative approach for the treatment of allergic rhinitis.
- Subjects :
- 0301 basic medicine
Male
Cancer Research
Receptors, CCR3
Down-Regulation
Bone Marrow Cells
Biology
Biochemistry
Small hairpin RNA
03 medical and health sciences
Mice
0302 clinical medicine
Downregulation and upregulation
Western blot
murine CC chemokine receptor 3
RNA interference
Genetics
medicine
Gene silencing
short hairpin RNA
Animals
Humans
eosinophil
RNA, Messenger
RNA, Small Interfering
Molecular Biology
Cell Proliferation
Messenger RNA
Mice, Inbred BALB C
medicine.diagnostic_test
Lentivirus
apoptosis
Articles
Eosinophil
Molecular biology
Eosinophils
030104 developmental biology
medicine.anatomical_structure
HEK293 Cells
Oncology
Terminal deoxynucleotidyl transferase
030220 oncology & carcinogenesis
Molecular Medicine
RNA Interference
Plasmids
Subjects
Details
- Language :
- English
- ISSN :
- 17913004 and 17912997
- Volume :
- 15
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Molecular Medicine Reports
- Accession number :
- edsair.doi.dedup.....adf6786a60ce7c9bd1683e640bdd1807