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In recent years, an increasing number of young women have been diagnosed with cancer, including some nulliparous women. Therefore, many young patients with early-stage cancer desire to preserve fertility after cytotoxic oncological treatments. It is important to develop a multidisciplinary approach to achieve the best outcomes for each patient. On the other hand, there has been a sharp increase in microRNAs (miRNAs) as potential biomarkers for the diagnosis, prognosis, and evaluation of treatment efficacy of several diseases. MiR-543 has been reported to affect the pathogenesis and progression of diseases via complex mechanisms. Understanding the regulatory role of miR-543 may aid comprehension of the pathogenesis and treatment of a broad range of diseases. Therefore, we provide an overview of the biogenesis, function, and role of miR-543 in various systems. These results shed light on the anticancer and endometrial protection role of miR-543 in young patients with gynecologic tumors and highlight the clinical potential of miR-543-based applications and related challenges.