Polydopamine (coded as PDA) is reported to be very adhesive and reactive due to the attached functional groups, such as amine and hydroxyl groups. In this work, taking advantage of the condensation between Si-OH of the hydroxylated alkyltrichlorosilane (ATS) and C-OH on PDA surface, ATS molecules with different chain carbon number of 10, 14 and 18 were grafted onto the 3-aminopropyl triethoxysilane (APTES)-PDA dual-layer film, which was composed of PDA outerlayer and APTES underlayer, on Si substrate. Thus, hydrophobic trilayer films coded as APTES-PDA-ATS were prepared successfully. In order to reveal the dependence of the tribological behaviors upon the microstructures of the film, tribological experiments were conducted on an atomic force microscope and a ball-on-plate tribometer. Experimental results showed that the (micro-) friction reducing (characterized by a parameter of relative friction coefficient or friction coefficient, RFC or FC) and macro-wear resisting (characterized by anti-wear life) behaviors were related with the chain length of outerlayer and the PDA thickness. Specifically, on one hand, as the chain length increases, RFC/FC decreased and macro anti-wear life lengthened; on the other hand, as PDA gets thicker, RFC increased and the anti-wear life enhanced.