Objective: This in vitro study evaluated the microleakage of Class V cavities restored with a resin composite and different adhesive systems after occlusal loading. Methods & Materials: Standardized box-shaped Class V cavities were prepared on the buccal side of 135 non-carious human premolars. The teeth were randomly divided into three groups of 45 premolars each and restored as follows: Group 1-two-step total-etch adhesive (Single Bond, 3M) + resin composite (Supreme, 3M ESPE); Group 2-two-step self-etch adhesive (Clearfil SE, Kuraray) + resin composite and Group 3-one-step self-etch adhesive (Xeno III, Dentsply) + resin composite. The restorations were finished with aluminum oxide discs (Sof-Lex, 3M). Fifteen teeth in each group received 10,000 × 100 N and 250 N occlusal loads, respectively, and the remaining 15 teeth served as the control. The premolars were immersed in 2% methylene blue for 24 hours. The dye penetration was examined under a stereo-microscope, and the results were statistically analyzed by Kruskal Wallis, Mann-Whitney U and Wilcoxon Signed Rank tests to determine differences between the groups. Results: Gingival margins showed more dye penetration than occlusal margins in all the tested groups (p<0.05). In all the tested adhesive systerns, 100 N occlusal loading did not change dye penetration; however, Groups 1 and 2 exhibited better marginal sealing than Group 3 at the enamel margins under 250 N occlusal loading. Conclusion: Within the limitations of this in vitro study, it may be concluded that enamel margins provided better marginal sealing than dentin/cementum margins and the two-step self-etch adhesive exhibited better marginal sealing than an all-in-one adhesive at the enamel margins under 250 N occlusal loading.