Our study includes inhibition of glycolysis with HK2 target and inhibition of HER2 pathway, which is highly expressed in many cancer types, with HER2 target. Reduced survival, negative metastasis, and induced apoptosis are expected due to the combined inhibition of these two targets. Trastuzumab was used as the HER2 inhibitor, while lonidamine was used as the HK2 inhibitor. Trastuzumab is an antibody designed to selectively bind to an antigen called human epidermal growth factor 2 (HER2). Lonidamine is a derivative of indazole-3-carboxylic acid, which is known to inhibit aerobic glycolysis in cancer cells. Lonidamine was delivered to cells in hybrid nanoparticles to increase the effect of lonidamine and make it target-specific. The combination of trastuzumab and Lonidamine-loaded hybrid nanoparticles yielded positive results. Increased apoptosis in SKBR3 cells was demonstrated by Annexin V, DNA fragmentation analysis, and gene-level BAX/BCL-2 ratio. In addition, increased VDAC expression in the combination group was detected by western blot analysis. The effect of the combination of trastuzumab and lonidamine loaded nanoparticles on metastasis was determined by scratch assay and MMP-2 gene expression analyses. According to the results, the combination group gave a high significance wound area compared to all other groups. In addition, MMP-2 gene expression showed significant inhibition in the combination group compared to the control. According to our results, the combination of trastuzumab and lonidamine-loaded hybrid nanoparticles created a synergistic effect. With this combined therapy, drug resistance can be prevented. It may also be beneficial to the chemotherapy process in HER2-positive breast cancer |