Abstract Replica symmetry breaking (RSB) has been widely recognized as a statistical analysis approach to understand the disorder and nonlinear interactions in complex systems ranging from atoms to the cosmic scale. However, it is challenging to analyze the nonlinear optical characteristics of random laser (RL) in disordered gain medium via RSB due to the lack of a general RSB-based statistical analysis framework. In this work, we report the tunable RSB in polymer fiber RL, where the effects of temperature and different structures on RSB are investigated experimentally and theoretically. It experimentally proves that RSB in RL is not robust, and disorder and temperature are responsible for tunable RSB in RL, which contributes to the improvement of the statistical analysis framework for investigating the optical principles of RL using RSB. And the finding of the tunable RSB allows to investigate the dynamical differences for various RL systems, which broadens the directions for the use of spin-glass theory to explore the physical mechanism of RL.