Background and Objective: It has recently been shown that the application of amniotic membrane conditioned medium is effective in cancer treatment. In this study, the effect of amniotic stem cells conditioned medium on the activity of Hsp90 and Cdk4 expression, were investigated in cancer cells.

Materials and Methods: Two cancer cell lines HeLa and MDA-MB-231 were treated with the supernatant amniotic membrane conditioned medium for 24 hours and then the viability of cancer cells was evaluated by MTT assay. Immunocytochemistry assay was used to investigate changes in Cdk4 expression in cancer cells after being treated with the amniotic membrane conditioned medium. RT-PCR analysis was carried out to study the expression of Hsp90α (aa1) gene before and after treatment.

Results: The results from the MTT assay indicated that the viability of cancer cells decreased after treatment with amniotic membrane conditioned medium. The outcomes of immunocytochemistry showed a decrease in Cdk4 expression in treated cancer cells. Evaluation of Hsp90α (aa1) expression with RT-PCR analysis for cancer cells treated with amniotic membrane conditioned medium revealed a reduction in the expression of the Hsp90α (aa1) gene.

Conclusion: Based on the results, treatment of cancer cells with amniotic membrane conditioned medium, in addition to the reduction of cancer cell viability, leads to the induction of apoptosis through the inhibition of Hsp90 and Cdk4. According to the results, it seems that the inhibition of Hsp90 could be the possible mechanism behind the anticancer properties of amniotic membrane.