Transcription factor MYB activates CTSF to affect gastric cancer cell proliferation and stemness through glutamine metabolism
Objective To investigate how the transcription factor MYB activates cathepsin F(CTSF)and affects gastric cancer cell proliferation and stemness through glutamine metabolism and to explore the underlying mechanisms.Methods We used the TCGA and KnockTF databases to analyze CTSFexpression in gastric cancer tissues and predict MYB as the transcription factor of CTSFand analyze MYB expression in gastric cancer,respectively.We performed dual-luciferase and ChIP assays to verify this binding relationship.We assessed CTSFand MYB expression using real-time PCR.We investigated cell proliferation using CCK-8 and colony formation assays.The cell spheroidal ability was detected using a sphere formation assay.We examined stem cell surface marker(OCT4,NANOG,and SOX2)and glutamine transporter(SLC1A5)expression using Western blotting.We used corresponding kits to measure the glutamine,glu-tamic acid,and α-ketoglutaric acid content as well as the NADPH/NADP+ratio,GSH/GSSG ratio,and aspartic and oxaloacetic acid levels.Results We detected low CTSFexpression levels in gastric cancer tissues and cells.CTSFoverexpression could inhibit gastric cancer cell stemness and proliferation.CTSFknockdown significantly increased the glutamine,glutamate acid,andα-ketoglutaric acid contents,NADPH/NADP+and GSH/GSSG ratios,and aspartic and oxaloacetic acid levels,and promoted cell proliferation and stemness.MYB was highly expressed in gastric cancer tissues and cells.Our bioinformatic prediction combined with ChIP and dual-luciferase experiments confirmed MYB as a transcription factor for CTSF.Conclusion The results of our study indicated that the transcription factor MYB acti-vated CTSFto promote gastric cancer cell proliferation and stemness through glutamine metabolism.
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