Synthetic lethality (SL) is a lethal phenomenon with an important role in cancer treatment. This study was conducted to analyze the hotspots and frontiers in SL research. The Web of Science Core Collection (WOSCC) was used to identify the 100 top-cited articles related to SL research. Additionally, wee1 inhibitors combined with erastin were used to determine the effectiveness of SL in vitro and in vivo. Relevant articles were published mainly from 2009 to 2021, reaching a peak in 2020; articles published in 2010 were most frequently cited among the 100-top cited papers. Most studies (54%) were performed in the United States. Articles in Nature Chemical Biology were cited more frequently than articles in other journals, whereas Nature published the largest number of reports on SL. Among the 88 corresponding authors, CJ Lord was the most productive. Overlay visualization of keyword analysis revealed that the hotspots in SL research were PARP inhibitors, BRCA mutations, DNA damage repair, and carcinogenesis. CRISPR, ferroptosis, wee1, double-strand break (dsb) repair, myc, immunotherapy, and replication stress are emerging topics in SL research, whereas ovarian cancer, prostate stress, acute myeloid leukemia, and other topics have been used as disease models in recent years. The application and therapeutic strategy of SL in cancer is an emerging trend. Significantly, experiments verified that the wee1 inhibitor AZD1775 and ferroptosis activator erastin have synergistic effects on ovarian cancer in vitro and in vivo. Combining bibliometric analysis with experimental verification is a useful approach for SL research.