High temperatures are a significant stress factor for plants. In fact, many biochemical reactions involved in growth and development are sensitive to temperature. In particular, heat stress (HS) represents a severe issue for plant productivity and strategies to obtain high yields under this condition are important goals in agriculture. While selenium (Se) is a nutrient for humans and animals, its role as a plant micronutrient is still questioned. Se can prevent several abiotic stresses (drought, heat, UV, salinity, heavy metals), but the action mechanisms are poorly understood. Se seems to regulate reactive oxygen species (ROS) and to inhibit heavy metals transport. In addition, it has been demonstrated that Se is essential for a correct integrity of cell membranes and chloroplasts, especially the photosynthetic apparatus. Previous results showed that in tobacco (Nicotiana tabacum cv. Bright-Yellow 2) cultures HS (5 min at 50 °C) induced cell death with apoptotic features, accompanied by oxidative stress and changes in the levels of stress-related proteins. In this work we investigated the effect of Se on the responses induced by HS. The obtained results show that Se markedly reduces the effects of HS on cell vitality, cytoplasmic shrinkage, superoxide anion production, membrane lipids peroxidation, activity of caspase-3-like proteases, and the levels of some stress-related proteins (Hsp90, BiP, 14-3-3s, cytochrome c).