Tarantula cubensis venom (Theranekron®D6) is widely used in veterinary medicine as a drug with anti-tumor, wound healing, anti-inflammatory, antiphlogistic properties. The purpose of the study was to explore Theranekron®D6 (TD6)’s antibacterial activity and its impact on apoptotic cell death in human colon and liver cancer cell lines. TD6 showed a dose and time dependent cytotoxic effect at 12h and 24h in HT-29 and HUH7 cancer cell lines. The IC50 values of TD6 were calculated as 12.18 µg/mL and 25.10 µg/mL in HT-29 and HUH-7 cell lines at 24h, respectively. TD6 induced apoptosis in HT-29 and HUH-7 cell lines. In these cells exposed to TD6, while the BAX/BCL-2 ratio and CASP-3 mRNA level increased, the HSP90 mRNA and protein level decreased. Also, TD6 exhibited antimicrobial activities against Pseudomonas aeruginosa, Enterococcus faecalis, Klebsiella pneumonia, Staphylococcus aureus, Candida albicans and Candida utilis. Obtained results demonstrated that TD6 has great potentials as alternative therapeutic for cancer and infectious diseases as apoptosis inducer and antimicrobial agent.