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Elicitors are used to increase <jats:italic>Ganoderma<\/jats:italic> bioactive composite production. This study aims to evaluate the antiproliferative activity of ethanolic extracts from mycelium of <jats:italic>Ganoderma tuberculosum (G. tuberculosum)<\/jats:italic> grown in a liquid medium with vineyard pruning waste (VPW) extracts as elicitors. Ethanolic and aqueous VPW extracts contain resveratrol dimer 4, resveratrol tetramer 1, and naringenin, while toluene and chloroform extracts contain tetradecanoic acid, hexadecanoic acid, and octadecanoic acid. Polar and nonpolar extracts could be promising elicitors for increasing bioactive molecules. Catechin gallate showed the highest correlation (<jats:italic>r<\/jats:italic>\u2009=\u20090.66) with biomass. Mycelial ethanolic extracts of <jats:italic>G. tuberculosum<\/jats:italic> (native strain from the Sonoran Desert) and <jats:italic>Ganoderma lucidum<\/jats:italic> (<jats:italic>G. lucidum<\/jats:italic>) (control) were analyzed by ESI\u2010IT\u2010MS, and 27 molecules were identified for the two species. They showed antiproliferative activity against the A549 and C\u201033 A cell lines but not for ARPE\u201019. <jats:italic>G. tuberculosum<\/jats:italic> culture with VPW had quinic acid, ganodermenonol, ganoderic acid I (GA\u2010I), C2 (GA\u2010C2), and 20\u2010hydroxylucidenic acid P, among others. Molecular docking of ganodermenonol, GA\u2010I, and GA\u2010C2 demonstrates significant interaction with tumor necrotic factor (TNF\u2010<jats:italic>\u03b1<\/jats:italic>). These ethanolic extracts of <jats:italic>Ganoderma<\/jats:italic> are promising sources of bioactive triterpenoids. Their antiproliferative activity did not change between species or treatment. Likewise, the <jats:italic>G. tuberculosum<\/jats:italic> and <jats:italic>G. lucidum<\/jats:italic> extracts only affected cancer cell lines. 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