Objective <p>Chemoresistance, such as paclitaxel (PTX) resistance, has become a great obstacle in non-small cell lung cancer (NSCLC) treatment. The natural agent salidroside (SAL) has been shown to exert an antitumor effect on NSCLC. Nonetheless, it is unclear whether SAL can decrease the resistance of NSCLC to PTX.</p> Methods <p>PTX-resistant NSCLC cells (H1299/PTX and A549/PTX) were generated. Cell Counting Kit-8 (CCK-8) assay was used to detect cell viability. Colony formation assay and flow cytometry were utilized to assess cell proliferation and apoptosis, respectively. Immunofluorescence staining and TOP/FOP flash luciferase assay were employed to estimate β-catenin activation. Western blotting was implemented to estimate the protein levels of apoptosis-, proliferation-, and Wnt/β-catenin signaling-associated markers. A xenograft mouse model was established to investigate the impact of SAL on PTX resistance in vivo.</p> Results <p>SAL increased PTX-induced suppression of proliferation and promoted apoptosis in PTX-resistant NSCLC cells. SAL blocked the Wnt/β-catenin signaling in A549/PTX cells and in tumor-bearing mice. Activating Wnt/β-catenin signaling reversed the SAL-mediated increase in the sensitivity of NSCLC cells to PTX. SAL attenuated PTX resistance in NSCLC in the xenograft mouse model.</p> Conclusion <p>SAL enhances the sensitivity of NSCLC cells to PTX by blocking the Wnt/β-catenin signal transduction.</p>