FASN and palmitate modifies H3K methylation and acetylation in Ut-LMS.

(A-B) Immunoblot showing FASN expression in SK-LMS-1 and FASN- or EV-transduced SK-UT-1 and Ut-LMS CRC cells. FASN was introduced by retroviral vector and positive clones (#2, 5, 6, 8) were selected by puromycin. Clone (#5) with the highest FASN expression was amplified. Non-transformed Ut-LMS cell line CRC-2 was established from patient tissue, and expresses FASN at a high level. (B) Time-dependent H3K alterations mediated by forced FASN overexpression in SK-UT-1. (C) H3K alterations mediated by FASN siRNA knockdown in SK-LMS-1. Scramble or FASN siRNA were transfected into high FASN expressing SK-LMS-1 cells and lysates were harvested at indicated time points. (D) Palmitate reproduces H3K modification in dose dependent manner in low FASN expressing parental SK-UT-1 cells. Cells were treated with palmitate at indicated dose (μM) for 48hr, and cell lysates were harvested for histone 3 detection. The images are representative of at least three experiments. Cropped blots are from gels run under same condition. FASN, fatty acid synthase; EV, empty vector; Ut, uterine; LMS, leiomyosarcoma. CRC, conditionally reprogrammed cells.