Cancer, one of the deadliest diseases worldwide, is projected to affect 30.2 million people by 2040. Among the various cancer types, rhabdomyosarcoma (RMS) is a unique tumor primarily impacting the muscular system of children. The current treatment for RMS has limited efficacy and numerous side effects, emphasizing the need for novel therapeutic approaches. This study investigates the potential treatment of the RMS cell line RD using extracts from five folklore-based medicinal plants in An Giang, Vietnam. The plants—Moringa oleifera Lam, Brucea javanica (L.) Merr., Eclipta prostrata (L.), Callisia fragrans (Lindl.) Woodson, and Zingiber zerumbet (L.) Smith—were extracted and fractionated using three solvents: ether, ethanol, and water. These fractions underwent phytochemical screening and cytotoxicity testing on the in-vitro RMS cell line RD. The results indicate that the ether fraction of Eclipta prostrata (L.) and the ether and ethanol fractions of Zingiber zerumbet (L.) Smith exhibit moderate cytotoxic effects on RD cell lines, with IC50 values of 37.08 ± 1.23 µg/mL, 23.15 ± 1.17 µg/mL, and 45.63 ± 2.39 µg/mL, respectively. These findings provide preliminary data for further in-depth research into the anticancer properties of these plants, which are widely grown in the South of Vietnam.