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Cystectomy is the primary treatment of muscle-invasive urothelial carcinoma, but it is associated with many complications and affects patients' quality of life. Chemotherapy is an alternative modality, but it may not give the expected response. This arouses the need for markers that help to predict the response to chemotherapy. Human epidermal growth factor receptor 2 (HER2-neu), S-phase kinase associated protein 2 (SKP2), and hypoxia-inducible factor-1 (HIF-1) regulate cell cycle progression, tumor adaptation to hypoxic environment, and response to chemotherapy.This study aimed at evaluation of HER2-neu, SKP2, and HIF-1 expressions in muscle-invasive urothelial carcinoma and investigated the association between their expressions and tumor response to chemotherapy.A total of 100 specimens from patients with nonmetastatic muscle-invasive urothelial carcinoma were collected at the pathology Department, and the selected patients received the treatment and follow-up at Clinical Oncology and Urology departments, Menoufia University. HER2-neu, SKP2, and HIF-1 expressions were evaluated using immunohistochemical techniques. The patients received chemotherapy followed by cystoscopic examination. Bladder biopsy was examined to determine tumor response.A significant association was found between partial tumor response to chemotherapy and HER2-neu, SKP2, and HIF-1 positive expression (P=0.004, 0.029, and 0.004). SKP2 expression was significantly associated with low apoptotic count and high mitotic one (P=0.008 and 0.01), whereas HIF-1 expression was significantly associated with necrosis (P=0.008). A statistically significant association was found between SKP2 and HR2-neu expression (P=0.018) and between SKP2 and HIF-1 expression as well (P=0.013).This study showed that evaluation of HER2-neu, SKP2, and HIF-1 expression can predict poor response to chemotherapy in muscle-invasive urothelial carcinoma and help in selecting patients who will benefit from chemotherapy. In addition, target therapy against these markers can be effective in treatment.