The Combinational Anti-Cancer Effects of Trastuzumab-Tarantula cubensis Alcoholic Extract on HER2-Overexpressing MDA-MB-453 Breast Cancer Cells


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Kılınç S., Günay N., Özgür A., Çankaya M., Kuzucu M.

9th Drug Chemistry Conference, Antalya, Turkey, 8 - 11 April 2021, pp.77

  • Publication Type: Conference Paper / Summary Text
  • City: Antalya
  • Country: Turkey
  • Page Numbers: pp.77
  • Erzincan Binali Yildirim University Affiliated: Yes

Abstract

Trastuzumab is a recombinant DNA-derived humanized monoclonal antibody that selectively inhibits HER2-positive breast cancer tumor formation as a single agent and in combination with other chemotherapeutics and natural products. Binding with high affinity to the extracellular domain of HER2 receptor, trastuzumab inhibits the proliferation of tumor cells that overexpress HER2 [1]. In order to increase therapeutic efficiency, administration of trastuzumab in combination with FDA approved drugs and natural products have been investigated in pre-clinical and clinical studies in cancer [1,2]. Venoms have been used in traditional medicine to treat various diseases for many years. Spider venom contains peptides and proteins which target a different receptor, ion channels, and enzymes. And these bioactive compounds exhibit cytotoxic properties to several cancer cells through apoptosis modulation, anti-proliferation, inhibition of enzymatic activities, and cell cycle alterations [3]. Tarantula cubensis alcoholic extract is prepared from the venom of spider T. cubensis. It is used as homeopathic agent which has shown anti-tumor, anti-inflammatory and antiphlogistic effects in veterinary medicine. It was also reported that T. cubensis alcoholic extract has high cytotoxic effects on various cancer types including breast cancer [4]. In this research, we investigated the synergistic anticancer effect of trastuzumab and T. cubensis alcoholic extract on HER2-positive human breast cancer cell (MDA-MB-453). The values of CI (Combination Index) were calculated using CompuSyn free software to identify synergistic effect of trastuzumab and T. cubensis alcoholic extract. Cell cycle, various stages of apoptosis and the presence of multiple caspases were detected with the Muse® Cell Cycle Kit, Muse® Annexin V & Dead Cell Kit and Muse®, MultiCaspase Kit, respectively. In addition, the expression of Bax, Bcl-2, Caspase-3, Caspase-9, Apaf-1 and β-catenin genes was determined by RT-qPCR, β-actin was used as a reference gene. Protein levels of Bax, Bcl-2, Caspase-3 were determined by ELISA. Additionally, Apaf-1, β-catenin, Caspase-9 expressions were detected by Western Blotting.