Proteomic analysis of mechanisms of hypoxia-induced apoptosis in trophoblastic cells.
Preeclampsia is often accompanied by hypoxia of the placenta and this condition induces apoptosis in trophoblastic cells. The aim of this study was to characterize global changes of apoptosis-related proteins induced by hypoxia in trophoblastic cells so as to clarify the mechanism of hypoxia-induced apoptosis by using the PoweBlot, an antibody-based Western array. Human choriocarcinoma cell line JAR was cultured for 24 hours under aerobic and hypoxic conditions. Hypoxia induced apoptosis accompanied by increased expression of Bcl-x, Caspase-3 and -9, Hsp70, PTEN, and Bag-1. Bad, pan-JNK/SAPK-1, Bcl-2, Bid, and Caspase-8 showed decreased expression. Hypoxia-induced apoptosis was increased with the transfection of a bag-1 antisense oligonucleotide. The bag-1 antisense oligonucleotide affected the expression of Bid, Bad, Bcl-2, JNK, and phosphorylated JNK, although expression of PTEN and Bcl-X did not change. Bag-1 may inhibit apoptosis by suppressing the expression of Bid and Bad. It may also enhance apoptosis by inhibiting the expression of Bcl-2 and by modulating phosphorylation of JNK. Both mitochondrial and stress-activated apoptosis pathways played important roles in the hypoxia induced cell death of trophoblastic cells. These findings will contribute to establish new approach to detect hypoxic stress of the placenta, which leads to preeclampsia and other hypoxia-related obstetrics complications.
Ishioka
Shin-ichi
S
Department of Obstetrics and Gynecology, Sapporo Medical University, School of Medicine, Chuou-ku, Sapporo, Japan. ishioka@sapmed.ac.jp
Ezaka
Yoshiaki
Y
Umemura
Kota
K
Hayashi
Takuhiro
T
Endo
Toshiaki
T
Saito
Tsuyoshi
T
D017209:Apoptosis; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000032:analysis / Q000502:physiology; D005260:Female; D006801:Humans; D048055:Mitogen-Activated Protein Kinase 8 / Q000378:metabolism; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D010766:Phosphorylation; D011247:Pregnancy; D040901:Proteomics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000032:analysis / Q000502:physiology; D014327:Trophoblasts / Q000378:metabolism / Q000473:pathology*
C091732:BCL2-associated athanogene 1 protein; D004268:DNA-Binding Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D014157:Transcription Factors; D048055:Mitogen-Activated Protein Kinase 8; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human