Antiproliferative and apoptotic effects of combined treatment of interferon alpha-2b (IFN) and vitamin e (vit E) on SK HEP-1 cells

LUCCI, ALVARO; CARLA GABRIELA COMANZO; VERA MARINA C.; LORENZETTI, FLORENCIA; FERRETTI, ANABELA C.; CEBALLOS, MARÍA P.; QUIROGA, ARIEL D.; ALVAREZ, MARÍA DE L.; CARRILLO, MARÍA C.

Rosario

Congreso; Reunión anual SAFIS 2019 ROSARIO; 2019

Our group demonstrated that IFN is an effective antitumoragent in the prevention and treatment of hepatocellularcarcinoma (HCC) given its apoptotic and antiproliferativeeffects. Besides, it was shown that vit E exerts inhibitoryeffects on liver cancer due to its apoptotic, antiangiogenicand antiproliferative activities. Previously, we observedthat combined treatment of IFN with vit E significantlydecreased cell viability, migration and invasion of humanHCC cell line SK HEP-1, in comparison with singletreatments.The aim of the present work is to deepen the study of theeffects of combining IFN and vit E, with particular focus onproliferation and apoptosis. Methods: SK-HEP 1 cells weretreated with 10000 U/I IFN and 25 uM δ-Tocotrienol (anisomer of vit E). Treatments were performed for 72 h usingsingle drugs (IFN-group and E-group) and theircombination (IFN-E-group). Also, a control group treatedwith drugs vehicles was included. We performed: a)annexin V-FITC assay to determine total apoptosis by flowcitometry, b) western blot studies to analyze theexpression of PCNA (marker of proliferation),proapoptotic Bax and antiapoptotic Bcl-Xl proteins, and c)dichlorofluorescein assay to evaluate reactive oxygenspecies (ROS) production. Results: IFN-E-group showed ahigher increase in total apoptosis (+520%*&), comparedwith monotherapies (IFN-group: +75%*; E-group: +90%*).Also, IFN-E-group showed a significant decrease in PCNAexpression (-35%*&) together with an increase in Baxprotein expression (+65 %*&) and a decrease in Bcl-Xlexpression (-85%*&), compared with monotherapies (IFNgroup: PCNA: -17%*, Bax: +23%*, Bcl-Xl: -55%*; E-group:PCNA: -20%*, Bax: +25%*, Bcl-Xl: -60%*). On the otherhand, IFN-E-group showed a significant increase in ROSproduction (+480%*&) compared with monodrugtherapies (IFN-group: +50%*, E-group: +150%*) (*p