| Du Jicong,Liu Ruling,Cheng Ying,Cai Jianming,Gao Fu,Liu Cong.Screening of the key miRNA downstream of TLR2 and validating the function of miR-21 in radioprotection[J].Chinese Journal of Radiological Medicine and Protection,2020,40(8):582-589 |
| Screening of the key miRNA downstream of TLR2 and validating the function of miR-21 in radioprotection |
| Received:April 01, 2020 |
| DOI：10.3760/cma.j.issn.0254-5098.2020.08.002 |
| KeyWords:Ionizing radiation Damage TLR2 RNA sequence miR-21 |
| FundProject:国家自然科学基金（81872559，81903260）；上海市青年科技英才扬帆计划（19YF1459100）；上海市卫计委科研课题青年项目（20174Y0173） |
| Author Name | Affiliation | E-mail | | Du Jicong | Department of Radiation Medicine, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China | | | Liu Ruling | Department of Radiation Medicine, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China | | | Cheng Ying | Department of Radiation Medicine, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China | | | Cai Jianming | Department of Radiation Medicine, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China | | | Gao Fu | Department of Radiation Medicine, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China | | | Liu Cong | Department of Radiation Medicine, Faculty of Naval Medicine, Naval Medical University, Shanghai 200433, China | victorliu20102020@163.com |
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| Abstract:: |
| Objective To screen the key miRNA downstream of TLR2 and explore the function of the miR-21. Methods Wild type (WT) and TLR2 KO mice were irradiated with 60Co γ-ray to compare their survivals. The downstream miRNAs of TLR2 signaling pathway were screened by RNA sequence in BMCs, and their expressions were verified by QT-PCR. Cell lines with overexpression or knockdown of a miRNA were established to evaluate the function of miRNA. Results The radiosensitivity of TLR2 KO mice was higher than that of TLR2 WT mice(χ2=4.490, 13.100, 7.928, P<0.05). The bone marrow transplantation experiment proved that the increased radiosensitivity of TLR2 KO mice was related to BMCs (χ2=4.291, P<0.05). A total of 55 differentially expressed genes were screened by RNA sequence ([log2 Fold Change]>0.95, Q<0.05), of which 28 were up-regulated and 27 were down-regulated. QT-PCR assay determined that miR-21 was down-regulated in BMCs of TLR2 KO (t=9.420, P<0.01) and MyD88 KO (t=10.700, P<0.01) mice. It was proved by QT-PCR that the expressions of IL-6 (t=13.790, P<0.05) and TNF-α (t=14.280, P<0.05) were increased in a TLR2 dependent manner after PAM3CSK4 stimulation. Overexpression of miR-21 promoted viability of EL4 cells (t=5.951, P<0.05) and NIH/3T3 cells (t=4.786, P<0.05) and reduced BMCs apoptosis in WT (t=4.842, P<0.05) and TLR2 KO (t=10.520, P<0.05) mice after radiation. Inhibition of miR-21 decreased the viability of EL4 cells (t=4.815, P<0.05) and NIH/3T3 cells (t=4.042, P<0.05). Conclusions miR-21 plays a key regulatory role in the process of TLR2 radioprotection, which may be related to the up-regulation of IL-6 and TNF-α. |
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