15-Deoxy-Δ12, 14-prostaglandin J2 activates PI3K-Akt signaling in human breast cancer cells through covalent modification of the tumor suppressor PTEN at cysteine 136. (28th June 2018) - Suh, Jinyoung
Abstract: 15-Deoxy-Δ 12, 14 -prostaglandin J2 (15d-PGJ2 ), one of the terminal products of cyclooxygenase-2-catalized arachidonic acid metabolism, has been shown to stimulate breast cancer cell proliferation and migration through Akt activation, but the underlying mechanisms remain poorly understood. In the present study, we investigated the effects of 15d-PGJ2 on the activity of PTEN, the inhibitor of the phosphoinositide 3-kinase (PI3K)-Akt axis, in human breast cancer (MCF-7) cells. Since the α, β-unsaturated carbonyl moiety in the cyclopentenone ring of 15d-PGJ2 is electrophilic, we hypothesized that 15d-PGJ2 -induced Akt phosphorylation might result from the covalent modification and subsequent inactivation of PTEN that has several critical cysteine residues. When treated to MCF-7 cells, 15d-PGJ2 bound to PTEN, and this was abolished in the presence of the thiol-reducing agent dithiothreitol. A mass spectrometric analysis by using recombinant and endogenous PTEN protein revealed that the cysteine 136 residue (Cys 136 ) of PTEN is covalently modified upon treatment with 15d-PGJ2 . Notably, the ability of 15d-PGJ2 to covalently bind to PTEN as well as to induce Akt phosphorylation was abolished in the cells expressing a mutant form of PTEN in which Cys 136 was replaced by serine (C136S-PTEN). The present study demonstrates for the first time that electrophilic 15d-PGJ2 directly binds to cysteine 136 of PTEN and provides new insight into PTEN loss in cancer progression associated with chronic inflammation. These observations suggest that 15d-PGJ2 can undergo nucleophilic addition to PTEN, presumably at Cys 136, thereby inactivating this tumor suppressor protein with concomitant Akt activation. Highlights: PTEN is inactivated through covalent modification by 15-deoxy-Δ 12, 14 -prostaglandin J2 in breast cancer MCF-7 cells. 15-Deoxy-Δ 12, 14 -prostaglandin J2 directly binds to the cysteine 136 residue of the recombinant PTEN. PTEN interaction with 15-deoxy-Δ 12, 14 -prostaglandin J2 stimulates Akt phosphorylation. 15-Deoxy-Δ 12, 14 -prostaglandin J2 stimulates growth of MDA-MB-231 cells when inoculated to the athymic nude mice.