Comparative Proteomics Analysis Reveals the Reversal Effect of Cryptotanshinone on Gefitinib-Resistant Cells in Epidermal Growth Factor Receptor-Mutant Lung Cancer

Abstract

Cryptotanshinone (CTS) is a lipophilic constituent of Salvia miltiorrhiza, with a broad-spectrum anticancer activity. We have observed that CTS enhances the efficacy of gefitinib in human lung cancer H1975 cells, yet little is known about its molecular mechanism. To explore how CTS enhances H1975 cell sensitivity to gefitinib, we figured out differential proteins of H1975 cells treated by gefitinib alone or in combination with CTS using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein–protein interaction (PPI) bioinformatic analyses of the differential proteins were performed. CTS enhanced H1975 cell sensitivity to gefitinib in vitro and in vivo, with 115 and 128 differential proteins identified, respectively. GO enrichment, KEGG analysis, and PPI network comprehensively demonstrated that CTS mainly impacted the redox process and fatty acid metabolism in H1975 cells. Moreover, three differential proteins, namely, catalase (CAT), heme oxygenase 1 (HMOX1), and stearoyl-CoA desaturase (SCD) were validated by RT-qPCR and Western blot. In conclusion, we used a proteomic method to study the mechanism of CTS enhancing gefitinib sensitivity in H1975 cells. Our finding reveals the potential protein targets of CTS in overcoming gefitinib resistance, which may be therapeutical targets in lung cancer.

Therapeutic effects of medicinal plants and their constituents on lung cancer, in vitro, in vivo and clinical evidence

“…Similarly, cryptotanshinone (CTS), a bioactive component of Salvia miltiorrhiza, was shown to sensitize gefitinib‐resistant EGFR‐mutant lung cancer cells by targeting catalase (CAT), heme oxygenase 1 (HMOX1) and stearoyl‐CoA desaturase (SCD). These proteins were identified through proteomic analysis and suggested to be potential therapeutic targets in lung cancers 248 …”

“…These proteins were identified through proteomic analysis and suggested to be potential therapeutic targets in lung cancers. 248 …”

Protein tyrosine kinase inhibitor resistance in malignant tumors: molecular mechanisms and future perspective

“…Cell membrane lipids remodeling led to activation of the EGFR/PI3K/AKT pathway, expedited cell proliferation and LPCAT1 expression, forming a positive feedback. 293 , 294 …”

“… 336 Additionally, cryptotanshinone (CTS) may inhibit the expression of the antioxidants such as catalase (CAT), HMOX1, and SCD and alleviate gefitinib resistance in NSCLC cells. 294 Since lipid peroxidation happens throughout ferroptosis, targeting lipid metabolism to induce ferroptosis or, even more broadly, targeting the tumor metabolic reprogramming may improve drug resistance. For instance, there are certain crosstalks between mannose metabolism and lipid metabolism.…”

“…Cell membrane lipids remodeling led to activation of the EGFR/PI3K/AKT pathway, expedited cell proliferation and LPCAT1 expression, forming a positive feedback. 293,294 There are also several amino acid metabolic pathways associated with drug resistance. First, phosphoglycerate dehydrogenase (PHGDH), a key enzyme of the serine synthesis pathway (SSP), is a major driver of sorafenib resistance in HCC.…”