Genistein Inhibits Pancreatic Cancer Cells Progression Through Regulation Of MiR-223and MiR-27a

Background and Objective: Pancreatic cancer (PC) is one of the most commonmalignances in digestive system. The5-year survival rate is less than6%in PC.Gemcitabine alone or in combination with other conventional therapeutics is thestandard of care for the treatment of advanced PC without any significant improvementin the overall survival of patients diagnosed with this deadly disease. It is known thatone of the reasons for this high mortality is due to drug resistance duringchemotherapeutic treatment. Therefore, it is pivotal to discover new potentialtherapeutic agents for the treatment of PC.(1)We investigated the effectof a naturalcompound genistein on cell growth,invasion and apoptosis in PC cell lines includingPANC-1, BxPC-3and AsPC-1.(2) We investigated whether a genistein coulddown-regulate miR-223, resulting in the inhibition of cell growth and invasion, andinduction of apoptosis in PC cells.(3)We investigated whether genistein coulddown-regulate miR-27a, resulting in the inhibition of cell growth and invasion, andinduction of apoptosis in PC cells.Methods:(1)We investigated the effect of genistein on cell growth, invasion, andapoptosis by MTT assay, Flow Cytometry (FCM), Wound-healing assay and Matrigelinvasion assay, respectively, in PANC-1, BxPC-3and AsPC-1cell lines.(2)miR-223and miR-27a expressions were examined by Real-time miRNA reversetranscriptase-PCR assay.(3)To explore the roles of miR-223and miR-27a in PC cells,we transfected PC cells with miRNA inhibitors. Western blotting analysis was used todetermine Fbw7and FOXO1protein expression. Results:1. Treatment of PC cells with genistein led to the inhibition of cell growth andinvasion, and induction of apoptosis in a does-dependent maner.2. Treatment of PC cells with genistein led to the down-regulation of miR-223,resulting in the up-regulation of Fbw7, which was correlated with inhibition of cellgrowth and invasion, and induction of apoptosis. Moreover, down-expression ofmiR-223inhibits cell growth and invasion, and induces apoptosis, with concomitantup-regulation of Fbw7, one of the target of miR-223.3. Genistein treatment of PC cells led to the down-regulation of miR-27a, resultingin the up-regulation of FOXO1. More importantly, we observed that down-expression ofmiR-27a inhibits cell growth and invasion, and induces apoptosis.Conclusions:1.Genistein inhibited cell growth, induced apoptosis, suppressed migration andinvasion in PANC-1, BxPC-3, and AsPC-1cells.2. Genistein inhibited cell growth, induced apoptosis, suppressed migration andinvasion in part mediated through down-regulation of miR-223and subsequentup-regulation of Fbw7in AsPC-1cell.3. Genistein played a tumor suppressor role in part through inhibition of miR-27ain PANC-1cell. Our results demonstrated that targeting miR-27a by genistein mayrepresent a potential strategy for treatment of pancreatic cancer.