*2.9. Gene Expression*

GES-1 and AGS cells were lysed with QIAzol Lysis Reagent (Qiagen, Hilden, Germany), RNA was purified using the miRNeasy Mini Kit (Qiagen, Hilden, Germany) and quantified by spectrophotometric analysis at 260 nm (NanoDrop ND-100, Thermo Fisher Scientific, Waltham, MA, USA). Primers for *IL-8*, *IL-6*, *PTGS1* (*COX-1*), *PTGS2* (*COX-2*) and *GAPDH* (housekeeping gene) genes Are reported in Table S1. *IL-8, IL-6, PTGS1* (*COX-1*), and *PTGS2* (*COX-2*) mRNA levels were investigated through SYBR Green method (iTaq Universal SYBR Green One-Step Kit, Bio-Rad Laboratories Srl, Segrate, Italy). The total RNA (10 ng/μL) from each sample was mixed with SYBR Green, corresponding primers and the reverse transcription enzyme, the real-time PCR was assessed using the CFX384TM Real-Time PCR Detection System (coupled to C1000TM Thermal Cycler; Bio-Rad Laboratories Srl, Segrate, Italy). The threshold values were set manually, the relative expression of each gene was calculated by normalizing the data on the basis of the housekeeping gene (GAPDH). The experiments were repeated, at least, three times.
