Background: Colorectal cancer (CRC) is one of the most common cancers in the world. Although surgical and screening techniques have vastly improved in the last 30 years, chemotherapeutics have not advanced sufficiently for successful treatment. Objective: The aim of this study is to investigate the microRNA (miRNA) expression changes and anticancer agent potential of Rubus Sanctus Schreb. root extract (RRE) on LoVo and HT-29 colorectal adenocarcinoma cell lines. Materials and Methods: LoVo and HT-29 CRC cell lines treated with different concentrations of RRE to find growth inhibitory effect with WST-1 assay. Fifty percent growth inhibition and 25% growth inhibition concentrations further evaluated with annexin V, total caspase, cell cycle, and migration assays. Real-time polymerase chain reaction was used to investigate the expression differences in miRNA after extract treatment. Results: Cell proliferation was reduced 77.98% in HT-29 cells after RRE treatment (P < 0.05). In the cell invasion analysis, RRE reduced invasion in both cell lines up to 75.56% (P < 0.05). In addition, RRE induced apoptosis in up to 98% of a cell population (P < 0.05). Similarly, pan-caspase activity increased to 97.6% and 87.2% in LoVo and HT-29 cell lines, respectively (P < 0.0001). After extract treatment, among the nine miRNAs evaluated, only miR-140 expression was significantly increased in both cell lines after RRE treatment (P < 0.05). Conclusion: Our data show for the first time that RRE has the capability to inhibit CRC cell proliferation and invasion and alter epigenetic mechanisms. Although further studies should be conducted on this topic, RRE is thought to be a potential candidate for the future studies regarding new therapy options.