Premium
RAGE Implications during DNA Double Strand Breaks in Trophoblast Cells
Author(s) -
Tsai K YF,
Hirschi K M,
Knowlton M N,
Mejia J F,
Hall P,
Davis T,
Reynolds P R,
Arroyo Juan A
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.802.81
Subject(s) - trophoblast , dna damage , apoptosis , microbiology and biotechnology , placenta , biology , andrology , dna , immunology , medicine , fetus , pregnancy , genetics
Intrauterine growth restriction (IUGR) is an obstetric pathology characterized by an increase in trophoblast apoptosis and decreased cell invasion. Trophoblasts are critical for a successful pregnancy and they mediate important steps such as implantation, immune protection of the fetus, maternal blood flow to the placenta, and delivery. Previous mouse studies in our laboratory showed increased trophoblast apoptosis, decreased cell invasion and increased placental receptors for advanced glycation end‐products (RAGE) during second hand smoke (SHS) treatment. More recently, a role for nuclear RAGE (nRAGE) has been implicated in DNA double strand break (DNA‐DSB) repair. We tested the hypothesis that RAGE is involved during DNA‐DSB repair in trophoblast cells. DSBs were induced in human trophoblasts (Bewo and Sw71) with cigarette smoke extract (CSE) or bleomycin (BLM). Assessment of nRAGE and g‐H2AX (involved in DNA‐DSB repair) was done by immunostaining. Invasion was measured in trophoblast cells after CSE and BLM treatment. Immunoblot was used to quantify nRAGE and g‐H2AX in cells. DNA degradation was used to determine DNA integrity. Increased Co‐expression of RAGE and y‐H2AX was observed in treated trophoblasts when compared to controls. Increases protein expression of RAGE and g‐H2AX was quantified by immunoblot in control and treated trophoblast cells . Trophoblast invasion was decreased 50% with 20ug/ml BLM treatment (p<0.0001) and 80% with 30ug/ml as compared to controls (p<0.0001). Invasion of trophoblast cells was decreased 92% (p<0.002) when treated with 0.5% CSE. Trophoblast DNA degradation was significantly detected following treatment with BLM or CSE. We conclude that CSE and BLM causes DNA‐DSBs in trophoblast cells and repair may plausibly be regulated by nRAGE and g‐H2AX. Confirmatory studies in the absence of RAGE are necessary in order to validate a possible link. DNA‐DSBs could be a factor in inhibited trophoblast invasion observed in compromised placenta. These studies provide a critical initial step in dissecting tobacco‐mediated IUGR progression. Support or Funding Information This work was supported by a grant from the Flight Attendant's Medical Research Institute (FAMRI, PRR and JAA) and a BYU Mentoring Environment Grant (JAA and PRR). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .