[Effects of exogenous suppressor of cytokine signaling on cell apoptosis in ectopic endometrial stromal cells].
OBJECTIVE
To explore the effects of exogenous suppressor of cytokine signaling 3 (SOCS3) in Janus Kinase/Signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathways and examine the relationship between exogenous SOCS3 and cell proliferation and apoptosis in ectopic endometrial stromal cells.


METHODS
Ectopic endometrial stromal cells from patients with histopathologically confirmed endometriosis were primarily cultured in vitro. Vimentin staining was used to identify the purity of endometrial stromal cells. Lentivirus containing SOCS3 and GFP (green fluorescent protein) recombinant (LV-SOCS3-GFP) or only GFP gene (LV-GFP) was used to transfect two groups of ectopic endometrial stromal cells from the same patient respectively. They were divided into experimental (EG) and negative (NC) control. The transfection efficiency of lentivirus was measured by GFP fluorescence expression under microscope. And the expressions of SOCS3, STAT3 and JAK2 mRNA and protein were analyzed by real-time polymerase chain reaction (PCR) and Western blot respectively. And phosphorylated JAK2 and STAT3 proteins (p-STAT3 and p-JAK2) were also analyzed by Western blot. Cell cycles and apoptosis were detected by flow cytometry.


RESULTS
The transfection efficiency of lentivirus was over 80% at 72 h post-transfection. Compared with NC, EG showed a higher expression of SOCS3 mRNA (P < 0.001). SOCS3 protein level of EG was higher than NC while p-STAT3 protein of EG obviously lower than NC. STAT3, JAK2 and p-JAK2 proteins showed no difference between two groups. The percentage of apoptotic cells in EG was higher remarkably than NC (22.0 ± 1.3)% vs (4.4 ± 1.3)%, P < 0.01. Moreover, EG cells showed a G0/G1 arrest compared with NC (87.0 ± 3.7)% vs (76.0 ± 3.7)%, P < 0.05.


CONCLUSIONS
Exogenous SOCS3 induces cell apoptosis and inhibits cell proliferation in ectopic endometrial stromal cells.
Department of Obstetrics & Gynecology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.
Min
M
Li
Department of Obstetrics & Gynecology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.
Fei
F
Ruan
Department of Obstetrics & Gynecology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.
Hong
H
Zhan
Department of Obstetrics & Gynecology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.
Xiangwei
X
Fei
Department of Obstetrics & Gynecology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.
Junyan
J
Ma
Department of Obstetrics & Gynecology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China. Email: linjun1088@163.com.
Jun
J
Lin
D017209:Apoptosis; D049109:Cell Proliferation; D002478:Cells, Cultured; D004715:Endometriosis / Q000378:metabolism* / Q000473:pathology*; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D017154:Stromal Cells / Q000378:metabolism*; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000235:genetics / Q000502:physiology*; D014162:Transfection
C106644:SOCS3 protein, human; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; C507924:JAK2 protein, human; D053614:Janus Kinase 2