CAJ | 학술논문

目的研究外源性细胞因子信号转导抑制因子3(SOCS3)对内异症患者异位子宫内膜间质细胞中Janus激酶/信号转导与转录活化因子3(JAK/STAT3)信号通路的调控及其对该细胞凋亡和增殖的影响.方法原代培养异位子宫内膜间质细胞;利用重组构建的SOCS3和GFP融合表达的慢病毒载体(LV-SOCS3-GFP)感染异位子宫内膜间质细胞,使其高表达SOCS3,设为实验组,同时将LV-GFP慢病毒载体感染同源细胞,设为对照组;实时荧光定量PCR检测各组细胞的SOCS3、JAK2、STAT3mRNA表达水平;蛋白免疫印迹检测各组细胞中SOCS3、JAK2、STAT3的蛋白表达情况,以及JAK2、STAT3蛋白的磷酸化水平;流式细胞仪检测外源性SOCS3对异位子宫内膜间质细胞凋亡及细胞周期的影响.结果慢病毒载体感染异位子宫内膜间质细胞后,实验组SOCS3 mRNA水平明显上调,与阴性对照组比较P＜0.01);实验组SOCS3蛋白表达显著上升,同时p-STAT3蛋白表达下调,而STAT3、JAK2及p-JAK2蛋白水平与对照组差异无统计学意义;实验组细胞凋亡比例(22.0±1.3)％,对照组(4.4±1.3)％,实验组细胞周期中G0/G1期比例(87.0±3.7)％,对照组(76.0±3.7)％,两指标比较均P ＜0.05.结论外源性SOCS3可促进异位子宫内膜间质细胞的凋亡并抑制细胞增殖.
목적 연구외원성세포인자신호전도억제인자3(SOCS3)대내이증환자이위자궁내막간질세포중Janus격매/신호전도여전록활화인자3(JAK/STAT3)신호통로적조공급기대해세포조망화증식적영향.방법 원대배양이위자궁내막간질세포;이용중조구건적SOCS3화GFP융합표체적만병독재체(LV-SOCS3-GFP)감염이위자궁내막간질세포,사기고표체SOCS3,설위실험조,동시장LV-GFP만병독재체감염동원세포,설위대조조;실시형광정량PCR검측각조세포적SOCS3、JAK2、STAT3mRNA표체수평;단백면역인적검측각조세포중SOCS3、JAK2、STAT3적단백표체정황,이급JAK2、STAT3단백적린산화수평;류식세포의검측외원성SOCS3대이위자궁내막간질세포조망급세포주기적영향.결과 만병독재체감염이위자궁내막간질세포후,실험조SOCS3 mRNA수평명현상조,여음성대조조비교P＜0.01);실험조SOCS3단백표체현저상승,동시p-STAT3단백표체하조,이STAT3、JAK2급p-JAK2단백수평여대조조차이무통계학의의;실험조세포조망비례(22.0±1.3)％,대조조(4.4±1.3)％,실험조세포주기중G0/G1기비례(87.0±3.7)％,대조조(76.0±3.7)％,량지표비교균P ＜0.05.결론 외원성SOCS3가촉진이위자궁내막간질세포적조망병억제세포증식.
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.Lentiviru 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.Conclusion Exogenous SOCS3 induces cell apoptosis and inhibits cell proliferation in ectopic endometrial stromal cells.