Objective To investigative the chemical components of Subergorgia suberosa collected from the South China Sea. Methods The Et₂O extract of S. Suberosa was purified by repeated column chromatography on silica gel, Sephadex LH-20, ODS, and semi-preparative RP-HPLC. The structures of obtained compounds were determined by spectroscopic analysis and comparison with previously reported data. The anti-tumor activity of 10 obtained 9,11-secosterols were tested with human lung adenocarcinoma A549 cell line and human osteo-sarcoma MG63 cell line; the antifungal activities of them were tested with Candida albicans and Aspergillus fumigatus in vitro. Results and Conclusion Ten 9,11-secosterols were isolated from Subergorgia suberosa collected from the South China Sea. Compounds 2 and 7 have been obtained for the first time from the genus of gorgonian Subergorgia. In vitro anti-tumor experiment showed that compounds 3-6 and 9 had different degrees of tumor inhibitory effects against cell lines A549 and MG63, with the IC₅₀ values being 6.31, 30.42, 8.33, 9.57 and 3.30 μg/mL for A549 cell line and 30.83, 21.32, 9.19, 28.01 and 8.62 μg/mL for MG63 cell line, respectively. The ten steroids showed no anti-fungi activity against Candida albicans or Aspergillus fumigatus in vitro.