Fluoride is widely distributed in nature, and at high concentrations, it targets the kidney and especially proximal tubule epithelial cells. Selenium is a typical trace element beneficial to humans, and the role of selenium in the prevention and treatment of fluoride-induced organ damage is an important research topic. The purpose of this study was to investigate the possible protective effects of selenium against fluoride-induced oxidative stress and apoptosis in rat renal tubular epithelial cells. We showed that the activity of antioxidant enzymes (superoxide dismutase and glutathione peroxidase) and total antioxidant capacity were significantly reduced in NaF-treated normal rat kidney cells (NRK-52E), whereas the levels of nitrogen monoxide (NO) and malondialdehyde (MDA) were significantly increased. Moreover, the number of apoptotic cells, mRNA expression of Bax, Bad, caspase-3, caspase-8, and caspase-9, and protein expression of Bax were elevated, while mitochondrial membrane potential and the protein expression of Bcl-2 were reduced. Compared with the NaF group, pretreatment with selenium enhanced the activity of antioxidant enzymes, mitochondrial membrane potential, and protein expression of Bcl-2, while the levels of NO and MDA, number of apoptotic cells, mRNA expression of Bax, Bad, caspase-3, caspase-8, and caspase-9, and protein expression of Bax were decreased. In conclusion, selenium exerted remarkable protective effect against NaF-induced oxidative stress and apoptosis and altered the expression of Bcl-2/caspase family.