Background and Objective: Though amniotic membrane has recently been introduced as a natural tissue with anti-cancer properties, there are inadequate studies about involved mechanisms. In this study, the effect of amniotic membrane culture supernatant on viability of cancer cells via applying different doses was evaluated. Apoptosis as a possible mechanism of inducing cancer cell death by amniotic membrane was also investigated. Materials and Methods: Amniotic membranes prepared from placentas in Caesarian sections which were high in epithelial level were cultured for 24 hours. Then, medium was collected and added in different volumes to the culture of cancer cells. After 24 hours, the viability of cancer cells was evaluated by the MTT assay. Apoptosis of cancer cells as a possible mechanism was assessed with the TUNEL assay. Analysis of data was performed using one-way Anova and post-Tukey test. Results: After the lapse of 24 hours, the outcome of the MTT assay revealed that supernatant of amniotic membrane significantly led to a decrease in the viability of cancer cells in comparison to the control group. Meanwhile, the TUNEL assay showed that induction of death in cancer cells was possibly through apoptosis which was dose-dependent. Conclusion: Owing to the dose-dependent effect of amniotic membrane on cancer cells and possible induction of apoptosis, more research is in demand to introduce amniotic membrane as a proper choice in the treatment of cancer.