The surface roughness and tribological properties of Cyclina sinensis shell cuticle were studied by using confocal laser scanning microscopy (CSLM) and UMT—2 micro-friction and wear testing machine respectively. The results showed that the descending order of the roughness of different parts of Cyclina sinensis shell cuticle was as follows: dorsal Cyclina sinensis shell, posterior part of Cyclina sinensis shell, abdominal Cyclina sinensis shell and anterior part of Cyclina sinensis shell. The transverse friction coefficient (TFC) was greater than the longitudinal friction coefficient (LFC) of the cuticle surface in the front and abdomen of Cyclina sinensis shell. However, TFC was less than LFC of that of posterior and back of Cyclina sinensis shell. This suggested that the tribological properties of Cyclina sinensis shell cuticle transverse and longitudinal had heterosexual characteristics. Comparing to the anhydrous test conditions, under the water lubrication conditions, TFC and LFC of the cuticle surface of the front of Cyclina sinensis shell both increased, TFC and LFC of the cuticle surface of the back of Cyclina sinensis shell both decreased, and TFC of the surface of the cuticle of the abdomen of Cyclina sinensis shell decreased, but LFC increased.