中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2014年
3期
191-195
,共5页
王文琦%邹艳芬%孙丽洲%张媛媛%左青
王文琦%鄒豔芬%孫麗洲%張媛媛%左青
왕문기%추염분%손려주%장원원%좌청
先兆子痫%细胞因子信号转导蛋白抑制因子%滋养层%细胞增殖%细胞运动
先兆子癇%細胞因子信號轉導蛋白抑製因子%滋養層%細胞增殖%細胞運動
선조자간%세포인자신호전도단백억제인자%자양층%세포증식%세포운동
Pre-eclampsia%Suppressor of cytokine signaling proteins%Trophoblasts%Cell proliferation%Cell movement
目的 探讨细胞因子信号转导抑制因子-3(suppressor of cytokine signaling-3,SOCS-3)基因在人子痫前期胎盘组织中的表达情况,及其对HTR-8/SVneo细胞增殖与迁移能力的影响. 方法 (1)选择2010年10月至2011年3月在南京医科大学第一附属医院住院分娩的重度子痫前期孕妇15例为子痫前期组,1 5例正常孕妇作为正常妊娠组.(2)体外培养的HTR-8/SVneo细胞分别转染SOCS-3小分子干扰RNA(实验组)和无意义的阴性对照小分子干扰RNA(阴性对照组).采用实时荧光定量逆转录 聚合酶链反应和Western印迹技术分别检测胎盘组织及体外培养细胞中SOCS-3 mRNA和蛋白表达水平;采用3-(4,5)-二甲基噻唑-(2,5)-二苯基溴化四氮唑蓝法检测细胞增殖能力;应用流式细胞仪检测细胞周期;采用Transwell小室实验检测细胞迁移能力.2组数据间比较采用两独立样本f检验. 结果 (1)子痫前期组胎盘组织SOCS-3 mRNA和蛋白表达水平分别为0.25±0.03和0.21±0.05,均低于正常妊娠组(0.71±0.08和0.75±0.12)(f值分别为15.94和14.29,P值均＜0.05).(2)小分子干扰RNA转染24 h后,实验组SOCS-3 mRNA水平低于阴性对照组(0.39±0.02与1.00±0.04,t=27.58,P＜0.05),蛋白水平也较低(0.003 7±0.001 4与1.514 9±0.035 7,t=73.35,P＜0.05).转染后,实验组细胞增殖能力降低,转染后48、72和96 h增殖能力分别为0.23±0.01、0.32±0.02和0.37±0.02,均低于阴性对照组(分别为0.39±0.02、0.55±0.04和0.86±0.04)(f值分别为2.60、6.64和42.44,P值均＜0.05).转染10d时实验组细胞克隆形成个数也低于阴性对照组[(116±15)个与(312±24)个,t=9.96,P＜0.05].转染48 h后,实验组G1/G0期细胞比例为(55.75±2.21)％,高于阴性对照组[(47.88±1.87)％](t=45.43,P＜0.05);S期细胞比例为(31.59±0.83)％,低于阴性对照组[(37.38±1.34)％](t=20.06,P＜0.05).转染48 h后,实验组穿膜细胞数为(93±11)个,低于阴性对照组[(167±17)个](t=21.36,P＜0.05). 结论 SOCS-3表达水平下降可能通过抑制滋养细胞的增殖和迁移能力,从而参与子痫前期的发生发展.
目的 探討細胞因子信號轉導抑製因子-3(suppressor of cytokine signaling-3,SOCS-3)基因在人子癇前期胎盤組織中的錶達情況,及其對HTR-8/SVneo細胞增殖與遷移能力的影響. 方法 (1)選擇2010年10月至2011年3月在南京醫科大學第一附屬醫院住院分娩的重度子癇前期孕婦15例為子癇前期組,1 5例正常孕婦作為正常妊娠組.(2)體外培養的HTR-8/SVneo細胞分彆轉染SOCS-3小分子榦擾RNA(實驗組)和無意義的陰性對照小分子榦擾RNA(陰性對照組).採用實時熒光定量逆轉錄 聚閤酶鏈反應和Western印跡技術分彆檢測胎盤組織及體外培養細胞中SOCS-3 mRNA和蛋白錶達水平;採用3-(4,5)-二甲基噻唑-(2,5)-二苯基溴化四氮唑藍法檢測細胞增殖能力;應用流式細胞儀檢測細胞週期;採用Transwell小室實驗檢測細胞遷移能力.2組數據間比較採用兩獨立樣本f檢驗. 結果 (1)子癇前期組胎盤組織SOCS-3 mRNA和蛋白錶達水平分彆為0.25±0.03和0.21±0.05,均低于正常妊娠組(0.71±0.08和0.75±0.12)(f值分彆為15.94和14.29,P值均＜0.05).(2)小分子榦擾RNA轉染24 h後,實驗組SOCS-3 mRNA水平低于陰性對照組(0.39±0.02與1.00±0.04,t=27.58,P＜0.05),蛋白水平也較低(0.003 7±0.001 4與1.514 9±0.035 7,t=73.35,P＜0.05).轉染後,實驗組細胞增殖能力降低,轉染後48、72和96 h增殖能力分彆為0.23±0.01、0.32±0.02和0.37±0.02,均低于陰性對照組(分彆為0.39±0.02、0.55±0.04和0.86±0.04)(f值分彆為2.60、6.64和42.44,P值均＜0.05).轉染10d時實驗組細胞剋隆形成箇數也低于陰性對照組[(116±15)箇與(312±24)箇,t=9.96,P＜0.05].轉染48 h後,實驗組G1/G0期細胞比例為(55.75±2.21)％,高于陰性對照組[(47.88±1.87)％](t=45.43,P＜0.05);S期細胞比例為(31.59±0.83)％,低于陰性對照組[(37.38±1.34)％](t=20.06,P＜0.05).轉染48 h後,實驗組穿膜細胞數為(93±11)箇,低于陰性對照組[(167±17)箇](t=21.36,P＜0.05). 結論 SOCS-3錶達水平下降可能通過抑製滋養細胞的增殖和遷移能力,從而參與子癇前期的髮生髮展.
목적 탐토세포인자신호전도억제인자-3(suppressor of cytokine signaling-3,SOCS-3)기인재인자간전기태반조직중적표체정황,급기대HTR-8/SVneo세포증식여천이능력적영향. 방법 (1)선택2010년10월지2011년3월재남경의과대학제일부속의원주원분면적중도자간전기잉부15례위자간전기조,1 5례정상잉부작위정상임신조.(2)체외배양적HTR-8/SVneo세포분별전염SOCS-3소분자간우RNA(실험조)화무의의적음성대조소분자간우RNA(음성대조조).채용실시형광정량역전록 취합매련반응화Western인적기술분별검측태반조직급체외배양세포중SOCS-3 mRNA화단백표체수평;채용3-(4,5)-이갑기새서-(2,5)-이분기추화사담서람법검측세포증식능력;응용류식세포의검측세포주기;채용Transwell소실실험검측세포천이능력.2조수거간비교채용량독립양본f검험. 결과 (1)자간전기조태반조직SOCS-3 mRNA화단백표체수평분별위0.25±0.03화0.21±0.05,균저우정상임신조(0.71±0.08화0.75±0.12)(f치분별위15.94화14.29,P치균＜0.05).(2)소분자간우RNA전염24 h후,실험조SOCS-3 mRNA수평저우음성대조조(0.39±0.02여1.00±0.04,t=27.58,P＜0.05),단백수평야교저(0.003 7±0.001 4여1.514 9±0.035 7,t=73.35,P＜0.05).전염후,실험조세포증식능력강저,전염후48、72화96 h증식능력분별위0.23±0.01、0.32±0.02화0.37±0.02,균저우음성대조조(분별위0.39±0.02、0.55±0.04화0.86±0.04)(f치분별위2.60、6.64화42.44,P치균＜0.05).전염10d시실험조세포극륭형성개수야저우음성대조조[(116±15)개여(312±24)개,t=9.96,P＜0.05].전염48 h후,실험조G1/G0기세포비례위(55.75±2.21)％,고우음성대조조[(47.88±1.87)％](t=45.43,P＜0.05);S기세포비례위(31.59±0.83)％,저우음성대조조[(37.38±1.34)％](t=20.06,P＜0.05).전염48 h후,실험조천막세포수위(93±11)개,저우음성대조조[(167±17)개](t=21.36,P＜0.05). 결론 SOCS-3표체수평하강가능통과억제자양세포적증식화천이능력,종이삼여자간전기적발생발전.
Objective To investigate the expression of suppressor of cytokine signaling-3 (SOCS-3) gene in placenta,its role in the pathogenesis of pre-eclampsia and its effect on proliferation and migration of HTR-8/SVneo cells.Methods Fifteen women with severe pre-eclampsia hospitalized in the First Affiliated Hospital of Nanjing Medical University from October 2010 to March 2011 and t 5 normal pregnant women during the same time period were investigated.Cultured HTR-8/SVneo cells were transfected with SOCS-3 specific small interfering RNA (siRNA) or negative siRNA as the controls.The expression of SOCS-3 mRNA and protein in placenta and these cells was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot.Cell proliferation was detected by methyl thiazolyl tetrazolium,cell cycle by flow cytometry and migration by the Transwell test.Two independent t tests were used for statistical analysis.Results The SOCS-3 mRNA and protein levels in the severe pre-eclampsia group were lower than those in the normal group (0.25±0.03 vs 0.71±0.08 and 0.21±0.05 vs 0.75±0.12,t=15.94 and 14.29,respectively,both P＜0.05).SOCS-3 mRNA and protein levels in the transfection group at 24 hours were lower than those in the negative control group (0.39±0.02 vs 1.00±0.04 and 0.003 7±0.001 4 vs 1.514 9±0.035 7,t=27.58 and 73.35,respectively,both P＜0.05).The integral absorbance values of cell proliferation in the transfection group at 48,72 and 96 hours after transfection were 0.23 ± 0.01,0.32±0.02 and 0.37± 0.02,respectively,which were lower than those in the negative control group (0.39± 0.02,0.55 ± 0.04 and 0.86± 0.04,t=2.60,6.64 and 42.44,respectively,all P＜0.05).The cell clonal formation was lower in the transfection group compared with the negative group (116± 15 vs 312±24,t=9.96,P＜0.05).The ratios of G1/G0 and S phase cells in the transfection group were (55.75±2.21) ％ and (31.59±0.83) ％,respectively,and were significantly different from those in the negative control group [(47.88± 1.87) ％ and (37.38± 1.34) ％,t=45.43 and 20.06,respectively,P＜0.05].After 48 hours,cell migration in the transfection group was lower than that in the negative control group (93 ± 11 vs 167± 17,t=21.36,P＜O.05).Conclusion SOCS-3 expression is probably involved in the pathogenesis of pre-eclampsia by being down-regulated and therefore impeding proliferation and migration of the trophoblast.
