Although the issue of left-behind children has received academic attention, little is known about the potential long-term effect. This study was conducted in China, aiming to explore the impact of left-behind experiences in childhood on entrepreneurial activities in adulthood. The study design was quantitative and we used secondary data with samples from the China Labor-force Dynamic Survey. To mitigate potential endogeneity problems, we adopted the conditional mixed process approach with instrumental variable. The empirical results show that left-behind experiences decreased the likelihood of adulthood entrepreneurship, which is supported by some robustness tests. This negative impact can be partially explained by disadvantages in human capital and social networks as well as risk aversion. Moreover, we found that the exogenous shocks from hukou transfer and WTO accession weaken the negative impact of left-behind experiences and lack of entrepreneurship can further explain the income penalization effect of left-behind experiences. Our study provides new evidence for the life course theory and reveals the long-term adverse effects of left-behind experiences in childhood. Also, we shed light on safeguarding the welfare of left-behind children and young people with left-behind experiences.