Cigarette smoking is a risk factor for carcinogenesis and the development of several types of cancer, including lung cancer. Cigarette smoke is considered to contain over 5,000 chemicals, including carcinogens. Heated tobacco products (HTPs) have been recently reported to reduce levels of toxic chemicals, such as nicotine and tobacco-specific N-nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), compared to burned cigarettes. We have previously reported that nicotine and NNK induce the proliferation of cancer stem cells (CSCs), which contribute to tumorigenicity, drug resistance and recurrence. To evaluated whether HTPs are involved in cancer development, we examined the effects of cigarette smoke extract (CSE) derived from HTPs on lung CSCs. We found that CSE induced the proliferation of lung CSCs and increased the expression levels of stem cell markers. Since CSCs exhibit epithelial-mesenchymal transition (EMT), we analyzed the expression of EMT markers. CSE induced expression of EMT markers, such as Twist and Snail. These results suggest that HTPs can promote CSC properties in vitro and may result in cancer development.