<p>Cancer is one of the leading causes of mortality worldwide. Development of new methods or improving the efficiency of already existing methods is essential in the successful treatment of this disease. Topotecan, a chemotherapeutic drug, has been used to inhibit various cancer types. However, chemotherapy resistance to this drug in cancer has impeded its maximum performance. miRNAs and other non-coding RNAs play crucial roles in regulating this attribute. In this systematic review, we investigated the interaction mechanism between these molecules and Topotecan in the modulation of cancer sensitivity to this agent. This study was carried out according to PRISMA guidelines. PubMed, Scopus, and Web of Science databases were comprehensively searched, using our predefined search terms. Following a selective process based on strategic criteria, eleven studies were included in the analysis. Altered expression levels of non-coding RNAs, especially miRNAs, regulated the sensitivity of cancer cell lines and animal models, directly and indirectly, through affecting cascades of signaling molecules. This impact was recorded in a variety of cancer types, including retinoblastoma, renal cell carcinoma, colorectal cancer, cervical cancer, breast cancer, prostate cancer, and leukemia. The highlighted interactions potentially offer new opportunities for modifying therapeutic intervention utilizing chemotherapeutic agents.</p>