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Effects of C-myc gene silencing on interleukin-1β-induced rat chondrocyte cell proliferation, apoptosis and cytokine expression.
- Source :
-
Journal of bone and mineral metabolism [J Bone Miner Metab] 2018 May; Vol. 36 (3), pp. 286-296. Date of Electronic Publication: 2017 Jun 14. - Publication Year :
- 2018
-
Abstract
- This study explores the effects of C-myc gene silencing on cell proliferation, apoptosis and cytokine expression in interleukin (IL)-1β-induced rat chondrocytes. Primary chondrocytes were obtained from 40 Sprague-Dawley rats. For in vitro C-myc3-shRNA transfection, chondrocytes were assigned to a blank 1, model 1, IL-1β + C-myc3-shRNA, C-myc3-shRNA, (IL-1β + C-myc3-shRNA) + C-myc overexpression, C-myc3-shRNA + C-myc overexpression or IL-1β + C-myc-Con group. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect C-myc, PCNA and cyclin D1 mRNA and protein expression. Cell proliferation was analyzed via CCK-8 assay and cell cycle while apoptosis was measured through flow cytometry. ELISA was utilized to assess the levels of metallopeptidase 13 (MMP-13), IL-6 and tumor necrosis factor-α (TNF-α). Both the qRT-PCR and Western blotting results demonstrated that C-myc3-shRNA transfection inhibits C-myc expression and promotes PCNA and cyclin D1 expression. In comparison to the model 1 group, all groups except the (IL-1β + C-myc3-shRNA) + C-myc overexpression and IL-1β + C-myc-Con groups showed increases in cell proliferation and S phase cell count and decreases in G <subscript>0</subscript> /G <subscript>1</subscript> phase cell count, cell apoptosis and MMP-13, IL-6 and TNF-α levels. The model 1, C-myc3-shRNA and C-myc3-shRNA + C-myc overexpression groups displayed higher cell proliferation and S phase cell count and reduced G <subscript>0</subscript> /G <subscript>1</subscript> phase cell count, cell apoptosis and MMP-13, IL-6 and TNF-α levels than the IL-1β + C-myc3-shRNA group. In comparison to the model 1 and C-myc3-shRNA + C-myc overexpression groups, the C-myc3-shRNA group promoted cell proliferation and S phase cell counts but suppressed G <subscript>0</subscript> /G <subscript>1</subscript> phase cell count, cell apoptosis and MMP-13, IL-6 and TNF-α levels. In conclusion, the study demonstrates that C-myc gene silencing can promote cell proliferation and inhibit cell apoptosis and cytokine expression in IL-1β-induced rat chondrocytes.
- Subjects :
- Animals
Cell Proliferation drug effects
Cells, Cultured
Chondrocytes drug effects
Cyclin D1 genetics
Cyclin D1 metabolism
Male
Osteoarthritis genetics
Osteoarthritis pathology
Proliferating Cell Nuclear Antigen genetics
Proliferating Cell Nuclear Antigen metabolism
Proto-Oncogene Proteins c-myc genetics
Proto-Oncogene Proteins c-myc metabolism
RNA, Messenger genetics
RNA, Messenger metabolism
RNA, Small Interfering metabolism
Rats, Sprague-Dawley
Transfection
bcl-2-Associated X Protein metabolism
Apoptosis drug effects
Chondrocytes cytology
Chondrocytes metabolism
Gene Silencing
Interleukin-1beta pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1435-5604
- Volume :
- 36
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Journal of bone and mineral metabolism
- Publication Type :
- Academic Journal
- Accession number :
- 28616752
- Full Text :
- https://doi.org/10.1007/s00774-017-0845-4