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ORIGINAL ARTICLE
Year : 2019  |  Volume : 37  |  Issue : 3  |  Page : 139-146

Explore the action of MiRNA-21 on shikonin and epidermal growth factor in regulating the proliferation and Apoptosis of HaCaT Cell


1 Department of Dermatology and STD, Zhejiang Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang, China
2 Department of Chinese Medicine Surgery, Zhejiang Chinese Medical University, Hangzhou, China
3 Department of Dermatology, Zhejiang Chinese Medical University, Third Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine, Hangzhou, China
4 Department of Dermatology and STD, Zhejiang Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang; Department of Chinese Medicine Surgery, Zhejiang Chinese Medical University, Hangzhou, China

Correspondence Address:
Dr. Yi Cao
Department of Dermatology and STD, Zhejiang Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, Youdian Rd., 54th, Hangzhou, 310006
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ds.ds_39_18

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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.


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