Mechanism of Abnormal Chondrocyte Proliferation Induced by Piezo1-siRNA Exposed to Mechanical Stretch
| Autor: | Haining Zhang, Dawei Li, Ping Leng, Yi Sun, Chenkai Li, Huanshen Gao, Zhenghui Li |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Small interfering RNA Article Subject Kinesins General Biochemistry Genetics and Molecular Biology Chondrocyte Ion Channels Flow cytometry 03 medical and health sciences 0302 clinical medicine Chondrocytes Downregulation and upregulation Western blot Tubulin medicine Humans RNA Small Interfering Cells Cultured Cell Proliferation Messenger RNA General Immunology and Microbiology medicine.diagnostic_test Chemistry Reverse Transcriptase Polymerase Chain Reaction PIEZO1 Cell Cycle Lentivirus General Medicine Transfection Flow Cytometry Cell biology 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation Medicine Stress Mechanical 030217 neurology & neurosurgery Research Article |
| Zdroj: | BioMed Research International, Vol 2020 (2020) BioMed Research International |
| ISSN: | 2314-6141 2314-6133 |
| Popis: | Objective. To investigate the effect of small interfering RNA targeting mechanosensitive ion channel protein Piezo1 (Piezo1-siRNA) on abnormal chondrocyte proliferation exposed to mechanical stretch. Methods. Construct and screen effective Piezo1-siRNA sequences and explore an appropriate method to transfect lentiviral vector into chondrocytes exposed to mechanical stretch. Western blot and RT-PCR were used to detect the mRNA and protein expression of Piezo1, Kif18A, and β-tubulin, respectively. Flow cytometry was used to measure the changes in the chondrocyte cycle. The proliferation of chondrocyte was evaluated by cell counting kit-8. Results. According to the mRNA and protein expression of Piezo1, the effective siRNA sequence was successfully screened. Compared with the 0 h group, mechanical stretch upregulated the expression of Piezo1, Kif18A, and β-tubulin, resulting in chondrocyte cycle arrest and eventually inhibiting chondrocyte proliferation. Moreover, Piezo1-siRNA transfection effectively blocks this process and promotes the proliferation of chondrocyte. Conclusion. Piezo1-siRNA can reduce the inhibition of chondrocyte proliferation induced by mechanical stretch via downregulating the expression of Kif18A and inhibiting the depolymerization of microtubules. Piezo1-siRNA plays a protective role in chondrocytes, which provides a potential method for the treatment of OA under abnormal mechanical stimulation. |
| Databáze: | OpenAIRE |
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