Cadmium (Cd) is a harmful heavy metal that is highly toxic to plants and animals. Expansins are cell wall proteins inducing cell wall loosening and participate in all plant growth and development processes which are associated with cell wall modifications. We investigated lettuce’s expansin gene LsEXPA6 and found that LsEXPA6 overexpression Arabidopsis lines were much more resistant to cadmium stress. Our results revealed that the root system of the expa6 mutant was suppressed under cadmium stress, resulting in shorter plant height, reduced biomass, and a significant increase in cadmium content in the plants compared with wild-type plants, whereas LsEXPA6 overexpression lines had a well-developed root system and reduced cadmium accumulation in the roots and shoots of the plants. The above results indicated that overexpression of LsEXPA6 affected root development and reduced Cd absorption in Arabidopsis. In addition, the higher absorption capacity of nutrients, increased antioxidant enzymes activities, improved chlorophyll and photosynthetic function in the overexpression Arabidopsis plants, supported the Cd stress tolerance mechanism. Taken together, these results provided a new insight on the role of expansin proteins in the tolerance of plants to Cd stress by root cell elongation.