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. 2020 Jun 30;25(2):100–110. doi: 10.15430/JCP.2020.25.2.100

Figure 6. The cyclopentenone ring on 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) is critical for 15d-PGJ2-induced apoptosis in MCF10A-ras cells.

Figure 6

(A) The chemical structures of 15d-PGJ2 and its non-electrophilic 9,10-dihydro-15d-PGJ2. An asterisk indicates an electrophilic carbon. (B, C) MCF10A-ras cells were treated with 15d-PGJ2 (10 μM) or 9,10-dihydro-15d-PGJ2 (10 μM) for 24 hours. Reactive oxygen species was measured by 2’,7’-Dichlorodihydrofluorescein diacetate staining. (C) Nuclear extracts from MCF10A-ras cells were incubated with [γ-32P]-labeled oligonucleotides harboring the NF-κB consensus sequence. The DNA binding activity were measured by electrophoretic mobility shifty assay. (D) MCF10A-ras cells were treated with 15d-PGJ2 (10 μM) or 9,10-dihydro-15d-PGJ2 (10 μM) for 24 hours, and cytotoxicity was measured by the MTT assay. Bars represent mean ± SE of three independent assays. A significant difference in the relative viability between treated cells and the solvent control is indicated with P < 0.01. (E) MCF10A-ras cells were treated with dimethyl sulfoxide, 15d-PGJ2 (10 μM), or 9,10-dihydro-15d-PGJ2 (10 μM) for 24 hours to measure the sub-G0/G1 population and the Annexin V-FITC/propidium iodide positive cell population. (F) The expression of proteolytic cleavage products of caspase-7 and PARP and that of IKKβ was determined by Western blot analysis in the MCF10A-ras cells treated for 24 hours with 15d-PGJ2 (10 μM) or 9,10-dihydro-15d-PGJ2 (10 μM).