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<p>Although ovine puberty initiation has been previously studied, the mechanism by which the RNA-binding protein Lin28B affects this process has not been investigated. The present study aimed to investigate the effects of <i>Lin28B</i> overexpression on <i>let-7b</i>, <i>let-7g</i>, cell proliferation, and estrogen secretion in Dolang sheep ovine ovarian granulosa cells. In this study, a <i>Lin28B</i> vector was constructed and transfected into ovarian granulosa cells using liposomes. After 24, 48, and 72 h of overexpression, quantitative real-time PCR (qRT-PCR) was used for measuring <i>let-7b</i> and <i>let-7g</i> microRNA (miRNA) levels, and estrogen secretion was measured using the enzyme-linked immunosorbent assay (ELISA). A CCK-8 (Cell Counting Kit-8) kit was used for evaluating cell viability and proliferation in response to <i>Lin28B</i> overexpression at 24 h. The results showed that the expression of <i>let-7b</i> and <i>let-7g</i> decreased significantly after <i>Lin28B</i> overexpression, and the difference was consistent over different periods. The result of ELISA showed that estradiol (E2) levels significantly increased following <i>Lin28B</i> overexpression. Additionally, <i>Lin28B</i> overexpression significantly increased the cell viability and proliferation. Therefore, the <i>Lin28B</i>–let-7 family axis may play a key role in the initiation of female ovine puberty.</p>