Objective  To verify the regulation of autophagy related molecules by hypoxia-inducible factor-1α (HIF-1α) through observation in vitro. Methods  Changes of expressions of HIF-1α and autophagy related molecules after human epithelial ovarian cancer cell lines being transfected by plasmids or intervened by drugs under normoxic or hypoxic culture condition were detected by Western blotting and immunofluorescence. Results  After being transfected by HIF-1α plasmids, HIF-1α was over-expressed, the expression of Beclin 1 increased, and LC3-Ⅰ converted to LC3-Ⅱ under the normoxic condition. After knockout of HIF-1α under the hypoxic condition, the expression of Beclin 1 decreased and LC3-Ⅱ induced by hypoxia was inhibited. Camptothecin drug NSC606985 inhibited the autophagy level of HIF-1α of high metastatic ovarian cancer cell lines. Conclusion  The hypoxic microenvironment can activate HIF-1α and induces the autophagy of epithelial ovarian cells. NSC606985 can regulate HIF-1α at nanomolar concentrations, but the effects on different cell types are different. The regulation may be positive or negative.