Differential expression of miR-672-5p and miR-146a-5p in osteoblasts in rats after steroid intervention
| Autor: | Yueguang Fan, Wenjun Feng, Yirong Zeng, Jianchun Zeng, Nan Sun, Pengfei Li, Jie Li |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
medicine.medical_specialty medicine.medical_treatment Biology Nucleic Acid Denaturation Real-Time Polymerase Chain Reaction Steroid 03 medical and health sciences 0302 clinical medicine ANGPTL4 Internal medicine microRNA Gene expression Genetics medicine Animals Gene Osteoblasts Gene Expression Profiling General Medicine Fold change Rats Gene expression profiling MicroRNAs 030104 developmental biology Endocrinology Real-time polymerase chain reaction 030220 oncology & carcinogenesis Steroids |
| Zdroj: | Gene. 591(1) |
| ISSN: | 1879-0038 |
| Popis: | Background Apoptosis of osteoblasts and osteocytes is one cause of steroid-induced osteonecrosis of the femoral head; however, the molecular mechanism of steroid affecting osteoblasts at the genetic level is unclear. The aim of the present work is to examine differential expression of osteoblasts in rats after steroid intervention and to verify expression by real-time polymerase chain reaction (RT-PCR). Methods Primary culture, passaging and identification of osteoblasts of SD neonatal rats were conducted; osteoblasts were divided into two groups, the control group, and the steroid group. Total RNA was extracted separately, and quality control was performed; by means of RNA labeling and microarray hybridization, data were collected and then standardized to ascertain differences in miRNA expression between the two groups. The gene expression spectrum was analyzed. Obvious differential expression of miR-672-5p and miR-146a-5p was verified by RT-PCR. Miranda, microcosm and mirdb bioinformatics software were used to predict target genes. Results Compared with the control group, morphologically, the osteoblasts in the steroid group were more irregular and showed various shapes. The number of miRNAs (fold change > 2) in the steroid group was six. Four miRNAs were upregulated and two miRNAs were downregulated. In particular, upregulated miR-672-5p expression and downregulated miR-146a-5p expression were significant. RT-PCR results showed that the 2−△△ CT value of miR-672-5p in the steroid group was 3.743-fold of that in the control group, and the 2−△△ CT value of miR-146a-5p in the steroid group was 0.322-fold of that in the control group. Angptl4, Ccdc51, Ssbp3 and RGD1306991 were predicted as the target gene of miR-672-5p, while Hrp12 was that of miR-146a-5p. Conclusion Expression profiles of miR-672-5p and miR-146a-5p had the most significant changes in the osteoblasts of rats with steroid intervention, which may provide a new viewpoint to pathogenesis of osteonecrosis of the femoral head. |
| Databáze: | OpenAIRE |
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