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Secalonic acid F (SAF) has been previously identified, however, little isknown about its cytotoxic activity and related cytotoxic mechanism. The aim ofthis study was to evaluate the cytotoxic activity of SAF isolated from a deepsea originated fungus Penicillium sp. F11 in HL60 cells and to analyze the differencesin protein expression of HL60 cells treated with SAF. The CCK-8 assay and Annexin V-FLUOS/PIassay indicated that SAF displayed dose- and time-dependent inhibition of HL60cell proliferation and induced apoptosis. Two-dimensional gel electrophoresis(2-DE) analysis of HL60 cells treated with SAF (4 µg/ml) revealed 10 differentiallyexpressed protein spots (P&lt;0.05), 5 upregulated and 5 downregulated. Threespots (1 downregulated and 2 upregulated) were identified as Rho GDP dissociationinhibitor 2 (RhoGDI 2) proteins by MALDI-TOF MS. Western blotting further demonstratedthe decreased abundance of full-length RhoGDI 2 together with the increased abundanceof caspase 3-cleaved product of RhoGDI 2. The caspase 3 inhibitor Ac-DEVD-CHOcould suppress the cytotoxic effect of SAF and significantly block the cleavageof RhoGDI 2. RhoGDI 2 is a cytosolic regulator of Rho GTPase and the caspase 3-cleavedproduct of RhoGDI 2 can advance progression of the apoptotic process. Our datashowed that SAF may modulate RhoGDI 2 levels in HL60 cells, thereby potentiallydisrupting cell signaling pathways important for HL60 cell function.

Differential proteomic analysis of HL60 cells treated with secalonic acid F reveals caspase 3-induced cleavage of Rho GDP dissociation inhibitor 2