Explore the action of MiRNA-21 on shikonin and epidermal growth factor in regulating the proliferation and Apoptosis of HaCaT Cell
| Autor: | Fengling Xing, Yang Xiaohong, Maocan Tao, Lili Ma, Yi Cao, Wei Ding, Hongbin Luo |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Cell
Dermatology Flow cytometry 030207 dermatology & venereal diseases 03 medical and health sciences 0302 clinical medicine Epidermal growth factor medicine lcsh:Dermatology MTT assay Viability assay shikonin medicine.diagnostic_test business.industry Cell growth Blood-heat syndrome psoriasis lcsh:RL1-803 Cell biology HaCaT medicine.anatomical_structure epidermal growth factor Apoptosis 030220 oncology & carcinogenesis business microRNA-21 |
| Zdroj: | Dermatologica Sinica, Vol 37, Iss 3, Pp 139-146 (2019) |
| ISSN: | 1027-8117 |
| Popis: | Purpose: The aim of the study is to investigate the effect of MicroRNA-21 (miR-21) and its interaction with epidermal growth factor (EGF) and shikonin on the proliferation, and apoptosis of HaCaT cell line. Materials and Methods: HaCaT cells were cultured under different concentrations of EGF and shikonin, and to calculate their optimal effect dosages. The transfection was performed using Lipofectamine2000, and then gene expression of miR-21 was detected by quantitative real-time polymerase chain reaction (RT-PCR). MTT assay and flow cytometry were applied to test cell proliferation and apoptosis. Western blot and RT-PCR were used to detect the proliferation (proliferating cell nuclear antigen [PCNA], NF-κB/IKKβ) and apoptosis (caspase-3/caspase-9, bcl-2) signals of HaCaT cell. Results: MTT assay showed that miR-21 mimic and EGF promoted, whereas, shikonin and miR-21 inhibitor inhibited cell viability of HaCaT cell. MiR-21 was upregulated by miR-21 mimic and EGF, while downregulated by shikonin and miR-21 inhibitor. Besides, EGF and miR-21 mimic promoted proliferation-associated signals (PCNA, NF-κB/IKKβ) expression, which were suppressed by shikonin and miR-21 inhibitor. Yet, shikonin and miR-21 inhibitor induced apoptosis-related signals (caspase-3/caspase-9, bcl-2) expression while reversed by EGF and miR-21 mimic which were confirmed by the result of flow cytometry. Conclusions: MiR-21 promotes the process of EGF-induced cell growth of HaCaT. The antagonized effect of shikonin in EGF-induced proliferation and apoptosis might be mediated by suppressing the expression of miR-21. |
| Databáze: | OpenAIRE |
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