Lipid metabolism plays an essential role in lung physiology—pulmonary environment is rich in specific lipid classes and enzymes involved in lipid synthesis, remodeling and degradation. Disruptions of lipid metabolism have been implicated in the pathogenesis of lung cancer, making the understanding of lipid signaling critical for the elucidation of underlying disease mechanisms and identification of potential therapeutic targets. It remains poorly understood how altered expression of different lipid hydrolases shape signaling, redox balancing and cellular responses such as proliferation or apoptosis of lung cancer cells. In this study, we investigated the impact of overexpression and knockout of the two key lipid hydrolases, adipose triglyceride lipase (ATGL) and monoacylglycerol lipase (MGL), on lipid signaling in lung cancer cell lines under nutrient-replete and lipid starved conditions. That was achieved by performing small-molecular thiols analyses and phospho-proteomics to map oxidative stress and the underlying signaling pathways. Lipase-dependent regulation of apoptosis was assessed using Annexin V assays. As a result, we report that stimulation of lipid hydrolysis causes cellular apoptosis under nutrient-replete as well as lipid starved condition and alters cellular redox status. Total glutathione levels, normalized to protein content, were reduced in lipase-overexpressing cells compared to controls under nutrient-replete conditions. Furthermore, lipid starvation decreased the ratio of reduced to oxidized glutathione relative to nutrient-replete conditions. In addition, we provide a comprehensive insight into phospho-proteome adaptation under differential expression of neutral lipases.