Water-pipe smoking (WPS), a common form of tobacco use, is particularly prevalent among young women in the Middle East. The smoke produced by WPS contains harmful substances similar to those found in cigarettes and is often associated with an increased risk of various cancers, including breast cancer. However, the specific genes affected by WPS and the mechanisms driving cancer initiation and progression, particularly in breast cancer, remain largely unknown. This study investigated the effects of chronic WPS exposure on normal human mammary epithelial cells. We analyzed the differential expression of genes using the NanoString nCounter PanCancer pathways panel, which includes 770 gene transcripts, and supplemented this with quantitative real-time polymerase chain reaction (PCR) analysis. The NanoString analysis revealed that 13 genes were significantly dysregulated due to WPS exposure. These genes are involved in various cellular processes, including signal transduction, cell cycle regulation, cell motility, proliferation, migration, invasion, and inflammation. Further, in silico analysis revealed that several of these genes were associated with breast cancer prognosis and were upregulated in breast cancer tissues compared to normal tissues. Notably, Kaplan-Meier survival analysis highlighted a strong correlation between dysregulation of WPS-related genes (MX1, CCL8, GNGT1, and MMP9) and relapse-free survival in breast cancer patients. The findings suggest that WPS exposure can significantly alter the expression of critical genes involved in breast cancer development and prognosis, suggesting its potential role in influencing breast cancer outcomes.
