Hybrid organic−inorganic halide perovskites (HOIPs) have received continued interest for their structure diversity and potential application in optoelectronic, solar cells, nonlinear optics (NLO), and ferroelectrics. Structural symmetry breaking induced by water molecules in single-crystal-tosingle-crystal (SCSC) transformations is beneficial to develop ferroelectrics or second-harmonic generation (SHG) materials. Along this line, a water-containing two-dimensional (2D) double perovskite, (C 6 H 16 N 2 ) 2 AgBiBr 8 •H 2 O (1), was prepared. Acentric 1 suffered a twice SCSC transformation when subjected to dehydration and rehydration, where the new centric (C 6 H 16 N 2 ) 2 AgBiBr 8 (2) and acentric (C 6 H 16 N 2 ) 2 AgBiBr 8 •0.5H 2 O (3) were generated. In contrast to the irreversible transformation from 1 to 2 (symmetry: P2 1 → Pmna), it is prominent that the reversible conversion of centric 2 to acentric 3 (symmetry: Pmna ↔ P2 1 2 1 2). The result validated the effect of guest water on inducing structural transformation and symmetry breaking of 2D perovskites, inspiring further explorations on water-involved 2D materials.