Background: The relationship between serum hormone levels and adolescent acne is not fully clarified. Objective: To determine the relationship between levels of androstenedione, dehydroepiandrosterone sulfate (DHEA-S), testosterone, estradiol and 17α-hydroxyprogesterone (17-OHP) with adolescent acne in Northeast China. Methods: A transversal study included 242 acne cases and 188 controls. All data were analyzed using SPSS version 17.0. Results: Androstenedione and testosterone levels were significantly higher (p < 0.0001) in the cases than in the control group. In males, the difference in 17-OHP levels was statistically significant (p < 0.0001), as well as between mild and severe acne cases (p = 0.002). The estradiol level was significantly different (p < 0.0001) between cases and controls in females. Conclusion: Higher androstenedione and testosterone levels are significant risk factors in the occurrence of adolescent acne. A higher 17-OHP level aggravates the severity of male adolescent acne, while a higher estradiol level protects females against the onset of adolescent acne.