Haemoglobin in pelvic fluid rescues Fallopian tube epithelial cells from reactive oxygen species stress and apoptosis

Fallopian tube fimbrial epithelium is considered to be the major site of origin of ovarian high‐grade serous carcinoma, with p53 loss being the earliest and universal change. We previously reported that reactive oxygen species (ROS) in the ovulatory follicular fluids (FFs) are mutagenic and cytotoxi...

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Published inThe Journal of pathology Vol. 240; no. 4; pp. 484 - 494
Main Authors Huang, Hsuan-Shun, Hsu, Che-Fang, Chu, Sung-Chao, Chen, Pao-Chu, Ding, Dah-Ching, Chang, Meng-Ya, Chu, Tang-Yuan
Format Journal Article
LanguageEnglish
Published Chichester, UK John Wiley & Sons, Ltd 01.12.2016
Wiley Subscription Services, Inc
Subjects
Animals
Apoptosis - physiology
Ascitic Fluid - metabolism
Cell Death - physiology
Cells, Cultured
DNA Breaks, Double-Stranded
DNA double-strand break
Epithelial Cells - cytology
Fallopian tube
Fallopian Tubes - cytology
Female
follicular fluid
haemoglobin
Hemoglobins - analysis
Hemoglobins - physiology
high-grade serous carcinoma
Humans
Menstruation - physiology
Mice, Knockout
NADPH Oxidases - physiology
Ovulation - physiology
Oxidative Stress - physiology
Reactive Oxygen Species - analysis
Reactive Oxygen Species - metabolism
high-grade serous carcinoma
haemoglobin
Fallopian tube
DNA double-strand break
follicular fluid
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Summary:Fallopian tube fimbrial epithelium is considered to be the major site of origin of ovarian high‐grade serous carcinoma, with p53 loss being the earliest and universal change. We previously reported that reactive oxygen species (ROS) in the ovulatory follicular fluids (FFs) are mutagenic and cytotoxic to fimbrial epithelial cells, which are bathed in the peritoneal fluid mixed with FFs. Here, we observed that ferryl haemoglobin (Hb), which was abundantly present in ovulatory FFs and pelvic peritoneal fluids, could rescue p53‐deficient immortalized fimbrial epithelial (FE25) cells and oviduct epithelial cells from Trp53‐null mice from lethal ovulatory ROS stress. Ferryl Hb and FF containing high Hb levels protected FE25 cells from apoptosis, mainly by consuming extracellular ROS and reducing NADPH oxidase‐mediated cell death. The remaining extracellular ROS could still induce DNA double‐strand breaks in the fimbrial epithelial cells. Our study revealed that ferryl Hb in peritoneal fluid rescued ROS‐stressed, DNA‐damaged fimbrial epithelial cells from death, and suggested that peritoneal blood from various sources may contribute to the ovulation‐induced transformation of Fallopian tube epithelium. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Bibliography:National Science Council - No. MOST103-2314-B-303-011-MY2
istex:D7B1B6D25CE811100A691C059A1077DA957920EA
ArticleID:PATH4807
Buddhist Tzu Chi General Hospital - No. TCRD105-24
Supplementary figure legendsFigure S1. Identification of the ROS-rescue factor in FFs. (A) FF aspirates had different rescue activities on H2O2 cytotoxicity on FE25 cells. The rescue activities of FFs were measured by treating FE25 cells with 500 μM H2O2 along with 3% FF (n = 17). Two types of FFs, rescue-high (H; n = 8) and rescue-low (L; n = 9), were identified. V: vehicle only (culture medium). (B) The flow chart of protein fractionation for identifying the rescue activity of FFs. Proteins in FFs were fractionated according to their molecular weight (f1: <3 kDa, f2: 3-10 kDa, and f3: >10 kDa) and each fraction was subjected to a rescue activity assay.Figure S2. MALDI-TOF MS analysis shows the distinct bends (130144-01 and 110043 in Figure 1D, arrow) in Rescue-H FFs are human haemoglobin β chain.Figure S3. Spectrophotometric analysis of ferryl Hb. (A) Spectrophotometry of the rescue-high FF (110144-01) revealed a 418 nm peak next to the 405 nm ferric Hb (methaemoglobin) , whereas the rescue-low FF (13013-09) did not show this peak (left panel). After treatment with 500 μM H2O2 for 1hr, ferric Hb (400 and 40 μg/ml) was oxidized to ferryl Hb and the spectrum shifted from 405 nm to 418 nm (right panel). (B) A standard curve was generated by serial dilution of ferryl Hb and absorbance at 418 nm. A regression R2 of 0.998 was established (right panel).Figure S4. Hb rescues H2O2-induced cell death. (A) Cell morphology shows cell death under lethal H2O2 (500 μM) treated (H) and survival by ferric Hb (Met) or ferryl Hb 100 μg/ml (H+Hb) treated. Scale bar: 50μm (B) Cell viability analysis shows that Hb (100 μg/ml ) added to FE25 cells with lethal H2O2-stressed (500 μM) can recapitulate rescue effect in five rescue-low FFs. **p<0.01 for each 0 μM H2O2 treated group.
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SourceType-Scholarly Journals-1
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ISSN:0022-3417
1096-9896
1096-9896
DOI:10.1002/path.4807