title: Disruption of paraoxonase 3 impairs proliferation and antioxidant defenses in human A549 cells and causes embryonic lethality in mice.
abstract: We had shown previously that paraoxonase 3 (PON3), a putative circulating antioxidant, was systemically upregulated in late-gestation rat, sheep, and human fetuses. Our overarching hypothesis is that preterm human infants are delivered with low levels of PON3 and that this contributes to a state of oxidative stress. We sought to determine whether absence of Pon3 was associated with reduced neonatal viability in mice and studied the offspring from crosses between Pon3(+/-) mice. The number of Pon3(-/-) animals at E10.5 and E17.5 was significantly lower than the expected 25% (9.3 and 7.9% respectively, P < 0.001). On the first day of postnatal life, this was reduced further (2.4%, significantly less than the proportion in fetal life, P = 0.04). Pon3(+/-) animals had lower body and placental weights than wild-type littermates at E17.5, an effect that was independent of the parent of origin of the mutant allele. We then studied the effect of PON3 knockdown in a human cell line, A549. Stable knockdown of PON3 using short-hairpin RNA reduced cell proliferation in 21% oxygen. We then studied the effect of transient knockdown of PON3 using short interfering RNA (siRNA) in the same cell line in low (2%) or ambient (21%) oxygen. Knockdown of PON3 using siRNA reduced total antioxidant capacity in 21% (P = 0.008) but not 2% oxygen. We conclude that the absence of Pon3 in mice resulted in increased rates of early fetal and neonatal death. Knockdown of PON3 in human cells reduced cell proliferation and total antioxidant capacity.
authors.affiliation: Department of Obstetrics and Gynaecology, University of Cambridge, Cambridge, UK.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Kempster
authors.forename: Gusztav
authors.initials: G
authors.lastname: Belteki
authors.forename: Diana
authors.initials: D
authors.lastname: Licence
authors.forename: D Stephen
authors.initials: DS
authors.lastname: Charnock-Jones
authors.forename: Gordon C S
authors.initials: GC
authors.lastname: Smith
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000975:Antioxidants / Q000378:metabolism; D043303:Aryldialkylphosphatase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D003433:Crosses, Genetic; D005260:Female; D005328:Fetal Viability; D055786:Gene Knockout Techniques; D020868:Gene Silencing; D005804:Genes, Lethal; D006579:Heterozygote; D006801:Humans; D008099:Liver / Q000196:embryology / Q000378:metabolism; D008168:Lung / Q000196:embryology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D018384:Oxidative Stress; D011247:Pregnancy; D034741:RNA, Small Interfering
chemical_list: D000975:Antioxidants; D034741:RNA, Small Interfering; D043303:Aryldialkylphosphatase; C113656:PON3 protein, human; C560866:PON3 protein, mouse