目的将调控细胞周期和增殖的E2F2基因导入成年哺乳动物耳蜗中,使其过表达,从而使耳蜗内的终末静止细胞重新进入细胞周期,向可分裂细胞转变,达到实现增加毛细胞数量的目的。方法将人来源的E2F2基因构建到腺病毒载体上,通过圆窗导入耳蜗;全耳蜗铺片、耳蜗切片观察E2F2过表达后耳蜗细胞的形态特点,用BrdU标记及ki67免疫组化方法检测其增殖情况。结果腺病毒携带的E2F2基因通过圆窗导入耳蜗后,能够在耳蜗毛细胞和支持细胞上表达。特别是在螺旋神经节细胞表达更强。转染E2F2基因后,耳蜗内的细胞BrdU标记呈阳性结果,且有ki67表达。结论过表达E2F2有可能使处于终末期的耳蜗细胞重新进入细胞周期,此方法为毛细胞再生研究提供了新的思路。
Objective To induce re-entry into cell cycles in cochlear terminal stationary cells via over expression of theE2F2 gene in adult mammals, for the purpose of increasing the number of hair cells. Methods Full-length human E2F2 gene was constructed into adenovirus vectors and introduced into the cochlea through the round window. Morphology of cochlear cells was studied using whole-mount and slice cochlear preparations. Cellular proliferation was detected with BrdU labeling and ki67 staining. Results E2F2 gene expression was seen in cochlear hair cells and supporting cells, as well as in spiral ganglion cells. Following E2F2 expression, there were also ki67and BrdU-positive cells in the cochlea. Conclusion Over-expression of E2F2 appears to allow end stage cells to re-enter the cell cycle. This technique may represent a promising new way of cell regeneration.