CAJ | 학술논문

目的：探讨miR-182诱导耳蜗前体细胞向毛细胞分化的作用及机制。方法从新生大鼠耳蜗中分离培养耳蜗前体细胞并用血清诱导分化，BrdU、Nestin免疫荧光染色鉴定细胞自我增殖能力；myosinⅦa和phalloidin免疫荧光染色鉴定其是否具有分化为毛细胞的潜能。分别利用miR-182 mimics和siRNA在新生大鼠耳蜗前体细胞中过表达和低表达miR-182，然后在细胞诱导分化7天后，用流式细胞仪检测分化细胞中myosinⅦa阳性细胞比例，用Western-blot检测支持细胞标志分子中Sox2的变化。结果使用miR-182 mimics进行过表达后，耳蜗前体细胞分化为myosinⅦa阳性表达的细胞比例显著高于对照组，使用miR-182 siRNA进行低表达后此比例显著低于对照组。Western-blot检测显示，Sox2在miR-182 mimics组较对照组降低，miR-182 siRNA组较对照组增加。结论过表达miRN182可以促进耳蜗前体细胞向毛细胞方向分化，Sox2可能是此过程中的靶基因。
목적：탐토miR-182유도이와전체세포향모세포분화적작용급궤제。방법종신생대서이와중분리배양이와전체세포병용혈청유도분화，BrdU、Nestin면역형광염색감정세포자아증식능력；myosinⅦa화phalloidin면역형광염색감정기시부구유분화위모세포적잠능。분별이용miR-182 mimics화siRNA재신생대서이와전체세포중과표체화저표체miR-182，연후재세포유도분화7천후，용류식세포의검측분화세포중myosinⅦa양성세포비례，용Western-blot검측지지세포표지분자중Sox2적변화。결과사용miR-182 mimics진행과표체후，이와전체세포분화위myosinⅦa양성표체적세포비례현저고우대조조，사용miR-182 siRNA진행저표체후차비례현저저우대조조。Western-blot검측현시，Sox2재miR-182 mimics조교대조조강저，miR-182 siRNA조교대조조증가。결론과표체miRN182가이촉진이와전체세포향모세포방향분화，Sox2가능시차과정중적파기인。
Objective To investigate the effects of miRNA182 in promoting differentiation of cochlear precursor cells. Methods Cochlear precursor cells were dissociated and cultured. The cultured cells were divided into three groups: (1) the control group;(2) the miRNA182 mimics group;and (3) the miRNA182 inhibiter group. To analyze the process of precursor cells differentiation, cells collected 7 days after culture were run through flow cytometry for myosinⅦ+cells. The expression of SOX2 was analyzed by Western-blotting. Results More myosinⅦ+ cells were observed when precursor cells were cultured with miRNA182 mimics than others. The expression of SOX2 was significantly suppressed in the miRNA182 mimics group. Conclusion Our findings suggest that miR-182 can promote in vitro cochlear progenitor cell differentiation. Sox2 may be one of the target genes during mir-182 induced process, providing new target for hair cell repairing.