Endometriosis is a serious human gynecological disorder of women of reproductive age. The present study was designed to explore the therapeutic implications of rhein in the management of endometriosis. The results showed that rhein significantly (p<0.05) inhibited the proliferation of endometrial stroma cells in a dose-dependent fashion. Besides, the rhein treated endometrial stroma cells showed significantly (p<0.05) lower migration and invasion, in vitro. Transwell and wound healing assays showed that rhein also suppressed the migration and invasion of the endometrial stroma cells. Rhein was shown to target miR-135 at the molecular level to exert its anti-proliferative effects against the human endometrial stroma cells. Conversely, overexpression of miR-135 could nullify the anti-proliferative effects of rhein. Taken together, the findings of the present study highlight the therapeutic utility of rhein against human endometriosis. However, more in vivo studies are required.