中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2012年
2期
129-133
,共5页
胡琳莉%钱坤%李海霞%孙莹璞%朱桂金
鬍琳莉%錢坤%李海霞%孫瑩璞%硃桂金
호림리%전곤%리해하%손형박%주계금
微RNAs%子宫内膜%间质细胞%蜕膜%微阵列分析%细胞周期
微RNAs%子宮內膜%間質細胞%蛻膜%微陣列分析%細胞週期
미RNAs%자궁내막%간질세포%세막%미진렬분석%세포주기
MicroRNAs%Endometrium%Stromal Cells%Decidua%Microarray analysis%Cell cycle
目的 研究在子宫内膜基质细胞( ESC)体外蜕膜化过程中的微小RNA (miRNA)差异表达谱及其细胞周期调控.方法 培养原代ESC,构建ESC体外蜕膜化模型(蜕膜化组),并进行形态学及催乳素水平验证,鉴定其构建成功；以未经蜕膜化处理的ECS作为对照(对照组).采用miRNA微阵列芯片技术筛选两组ESC中miRNA差异表达谱,利用实时荧光定量PCR技术对miRNA微阵列芯片检测结果进行验证.转染miR-222抑制物后,流式细胞仪检测两组体外蜕膜化ESC的细胞周期比例.结果 (1) miRNA微阵列芯片技术筛选结果显示,蜕膜化组与对照组中有49个ESCmiRNA表达水平具有显著差异,其中16个miRNA显著上调,33个miRNA显著下调.实时荧光PCR技术检测显示,hsa-miR-27b、30c、143、101、181b、29b、30d、507、23a、222、221的表达水平在蜕膜化组与对照组比较,差异均有统计学意义(P＜0.05).(2)对照组转染阴性对照-6-羧基荧光素(NC-FAM)后S期细胞比例为(6.2±0.7)％,明显低于转染前的(10.9±0.8)％(P＜0.05)；而G0/G1期细胞[(77.5±1.3)％]较转染前[(73.0±1.6)％]有升高趋势,但差异无统计学意义(P＞0.05).蜕膜化组转染后S期细胞比例[(3.3±0.6)％]较转染前[(7.8±0.9)％]明显减少(P＜0.05)；G0/G1期细胞比例[ (80.7±1.6)％]较转染前[(74.9±1.1)％]有升高趋势,但差异无统计学意义(P＞0.05).对照组和蜕膜化组G2/M期细胞比例转染前、后比较,差异均无统计学意义(P均＞0.05).结论 miRNA可能通过调控细胞周期进程参与ESC体外蜕膜化过程.
目的 研究在子宮內膜基質細胞( ESC)體外蛻膜化過程中的微小RNA (miRNA)差異錶達譜及其細胞週期調控.方法 培養原代ESC,構建ESC體外蛻膜化模型(蛻膜化組),併進行形態學及催乳素水平驗證,鑒定其構建成功；以未經蛻膜化處理的ECS作為對照(對照組).採用miRNA微陣列芯片技術篩選兩組ESC中miRNA差異錶達譜,利用實時熒光定量PCR技術對miRNA微陣列芯片檢測結果進行驗證.轉染miR-222抑製物後,流式細胞儀檢測兩組體外蛻膜化ESC的細胞週期比例.結果 (1) miRNA微陣列芯片技術篩選結果顯示,蛻膜化組與對照組中有49箇ESCmiRNA錶達水平具有顯著差異,其中16箇miRNA顯著上調,33箇miRNA顯著下調.實時熒光PCR技術檢測顯示,hsa-miR-27b、30c、143、101、181b、29b、30d、507、23a、222、221的錶達水平在蛻膜化組與對照組比較,差異均有統計學意義(P＜0.05).(2)對照組轉染陰性對照-6-羧基熒光素(NC-FAM)後S期細胞比例為(6.2±0.7)％,明顯低于轉染前的(10.9±0.8)％(P＜0.05)；而G0/G1期細胞[(77.5±1.3)％]較轉染前[(73.0±1.6)％]有升高趨勢,但差異無統計學意義(P＞0.05).蛻膜化組轉染後S期細胞比例[(3.3±0.6)％]較轉染前[(7.8±0.9)％]明顯減少(P＜0.05)；G0/G1期細胞比例[ (80.7±1.6)％]較轉染前[(74.9±1.1)％]有升高趨勢,但差異無統計學意義(P＞0.05).對照組和蛻膜化組G2/M期細胞比例轉染前、後比較,差異均無統計學意義(P均＞0.05).結論 miRNA可能通過調控細胞週期進程參與ESC體外蛻膜化過程.
목적 연구재자궁내막기질세포( ESC)체외세막화과정중적미소RNA (miRNA)차이표체보급기세포주기조공.방법 배양원대ESC,구건ESC체외세막화모형(세막화조),병진행형태학급최유소수평험증,감정기구건성공；이미경세막화처리적ECS작위대조(대조조).채용miRNA미진렬심편기술사선량조ESC중miRNA차이표체보,이용실시형광정량PCR기술대miRNA미진렬심편검측결과진행험증.전염miR-222억제물후,류식세포의검측량조체외세막화ESC적세포주기비례.결과 (1) miRNA미진렬심편기술사선결과현시,세막화조여대조조중유49개ESCmiRNA표체수평구유현저차이,기중16개miRNA현저상조,33개miRNA현저하조.실시형광PCR기술검측현시,hsa-miR-27b、30c、143、101、181b、29b、30d、507、23a、222、221적표체수평재세막화조여대조조비교,차이균유통계학의의(P＜0.05).(2)대조조전염음성대조-6-최기형광소(NC-FAM)후S기세포비례위(6.2±0.7)％,명현저우전염전적(10.9±0.8)％(P＜0.05)；이G0/G1기세포[(77.5±1.3)％]교전염전[(73.0±1.6)％]유승고추세,단차이무통계학의의(P＞0.05).세막화조전염후S기세포비례[(3.3±0.6)％]교전염전[(7.8±0.9)％]명현감소(P＜0.05)；G0/G1기세포비례[ (80.7±1.6)％]교전염전[(74.9±1.1)％]유승고추세,단차이무통계학의의(P＞0.05).대조조화세막화조G2/M기세포비례전염전、후비교,차이균무통계학의의(P균＞0.05).결론 miRNA가능통과조공세포주기진정삼여ESC체외세막화과정.
Objective To study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro.Methods ESC was induced decasualization in vitro and matched with non-decidualized cells as controls.The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR.Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it.Results (1) Between decidualized and undecidualized stromal cells,there were 49 miRNAs significantly different expression by microarray chip,including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b,30c,143,101,181 b,29b,30d,507,23 a,222,221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P ＜0.05).(2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC,ESC were cultured by FBS medium for 24 hours,the rate of transfection was 70％.ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours,the percentage of ESC at Sphase of (6.2 ± 0.7 ) ％ were significantly lower than ( 10.9 ± 0.8 ) ％ in control group ( P ＜ 0.05 ) ; the percentage of ESC at G0/G1 phase increased at transfection group [ (77.5 ± 1.3 ) ％ vs.(73.0 ± 1.6) ％ at control group ],but there was no significant difference (P ＞ 0.05 ).Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h,the percentage of ESC at S-phase was ( 3.3 ± 0.6) ％ in transfection group,which were significantly lower than (7.8 ± 0.9 ) ％ in control group ( P ＜ 0.05 ).The percentage of ESC at G0/G1 phase was ( 80.7 ± 1.6 ) ％ in transfection group and ( 74.9 ± 1.1 ) ％.In control group,which did not reached statistical difference ( P ＞ 0.05).Conclusion miRNA was involved in ESC decidual process in vitro by regulating cell cycle.
