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Purpose: Apatinib has been utilized in colon cancer therapies but its efficiency and molecular mechanism are not fully understood. Chemotherapy in combination with non-toxic compounds can be a strategy to reduce the recurrence of cancer. Consequently, this study was carried out to evaluate the effects of Apatinib and Piperine on colorectal cancer (CRC) cell line and their potential anti-cancerous mechanisms in vitro. Methods: The effects of Apatinib and Piperine on HCT-116 CRC cells were detected by assessing cell viability using MTT assay. The potential cytotoxic mechanisms of Apatinib and Piperine were investigated by evaluating the apoptosis-related gene (MDM-2) expression ratio using real-time PCR assay. Moreover, the glutathione peroxidase activity (GPX) and nitric oxide (NO) levels were assessed by colorimetric assays. Results: The proliferation rate of CRC cells decreased by increasing the concentrations of Piperine and Apatinib. When HCT-116 cells were treated with different concentrations of Apatinib in combination with Piperine, the synergistic effects were observed (Combination Index&lt;1). In HCT-116 cells treated with Apatinib or Piperine at the concentrations of 0.5×IC50 and 0.2×IC50, the MDM-2 gene expression was downregulated and NO levels increased compared to the untreated control cells and related single treatments. Furthermore, the cytotoxic effects of Apatinib increased when was combined with Piperine. In addition, GPX activity significantly decreased in combination treatment at 0.5×IC50 concentration of both agents. Conclusion: Apatinib in combination with Piperin could significantly inhibit the viability of CRC cells. These cytotoxic effects were induced by regulation of apoptosis-related gene and inhibition of antioxidant marker.

advanced pharmaceutical bulletin

Regulatory Effects of Apatinib in Combination with Piperine on MDM-2 Gene Expression, Glutathione Peroxidase Activity and Nitric Oxide level as Mechanisms of Cytotoxicity in Colorectal Cancer Cells