*Article* **Bothrops Jararaca Snake Venom Modulates Key Cancer-Related Proteins in Breast Tumor Cell Lines**

**Carolina Yukiko Kisaki 1, Stephanie Santos Suehiro Arcos 1, Fabio Montoni 1, Wellington da Silva Santos 1, Hamida Macêdo Calacina 1, Ismael Feitosa Lima 1, Daniela Cajado-Carvalho 1, Emer Suavinho Ferro 2, Milton Yutaka Nishiyama-Jr 1,\* and Leo Kei Iwai 1,\***


**Abstract:** Cancer is characterized by the development of abnormal cells that divide in an uncontrolled way and may spread into other tissues where they may infiltrate and destroy normal body tissue. Several previous reports have described biochemical anti-tumorigenic properties of crude snake venom or its components, including their capability of inhibiting cell proliferation and promoting cell death. However, to the best of our knowledge, there is no work describing cancer cell proteomic changes following treatment with snake venoms. In this work we describe the quantitative changes in proteomics of MCF7 and MDA-MB-231 breast tumor cell lines following treatment with *Bothrops jararaca* snake venom, as well as the functional implications of the proteomic changes. Cell lines were treated with sub-toxic doses at either 0.63 μg/mL (low) or 2.5 μg/mL (high) of *B. jararaca* venom for 24 h, conditions that cause no cell death per se. Proteomics analysis was conducted on a nano-scale liquid chromatography coupled on-line with mass spectrometry (nLC-MS/MS). More than 1000 proteins were identified and evaluated from each cell line treated with either the low or high dose of the snake venom. Protein profiling upon venom treatment showed differential expression of several proteins related to cancer cell metabolism, immune response, and inflammation. Among the identified proteins we highlight histone H3, SNX3, HEL-S-156an, MTCH2, RPS, MCC2, IGF2BP1, and GSTM3. These data suggest that sub-toxic doses of *B. jararaca* venom have potential to modulate cancer-development related protein targets in cancer cells. This work illustrates a novel biochemical strategy to identify therapeutic targets against cancer cell growth and survival.

**Keywords:** mass spectrometry; proteome; snake venom; *Bothrops jararaca*; breast cancer

**Key Contribution:** We describe that *B. jararaca* snake venom modulates specific protein pathways related to cancer cell growth and invasion, which could be a useful approach to identify novel therapeutic targets for cancer treatment.
