中国综合临床
中國綜閤臨床
중국종합림상
CLINICAL MEDICINE OF CHINA
2014年
9期
897-901
,共5页
细胞周期蛋白A%血管平滑肌细胞%起始识别复合物1
細胞週期蛋白A%血管平滑肌細胞%起始識彆複閤物1
세포주기단백A%혈관평활기세포%기시식별복합물1
Cyclin A%Vascular smooth muscle cells%Origin recognition complex 1
目的 探讨细胞周期蛋白A (Cyclin A)对培养大鼠血管平滑肌细胞(VSMCs)周期起始识别复合物1(ORC1)表达水平的影响.方法 采用组织块贴片法原代培养的大鼠胸主动脉血管平滑肌细胞,利用双胸苷使VSMCs达到细胞周期同步化,应用逆转录聚合酶链反应技术和流式细胞仪检测不同细胞周期VSMCs的ORC1和Cyclin A mRNA和蛋白表达.结果 经鉴定培养的细胞为VSMCs,采用血清饥饿法、双胸苷阻断法和秋水仙素阻抑法分别获得了处于不同细胞周期的同步化的VSMCs:G0/G1期(89.22 ±3.54)％,G1/S交界期(66.74±7.16)％,S期(63.24±4.06)％,G2/M期(51.64±11.18)％,各期比较差异有统计学意义(P均＜0.01).Cyclin A在静止状态对VSMCs的ORC1 mRNA和蛋白无影响,在G2/M期达到高峰[(52.133 3 ±2.122 1)％],与G1/S期[(10.9167±0.531 1)％]、S期[(7.656 7±0.412 4)％]比较,差异有统计学意义(P均＜0.01).ORC1在G2/M期达到最低值[(1.276 7 ±0.161 7)％],与G1/S期[(13.371 0±1.057 3)％]、S期[(3.043 3 ±0.538 0)％]比较,差异有统计学意义(P均＜0.01),可能原因是Cyclin A阻止ORC1结合在VSMCs染色质上.结论 CyclinA可能调控ORC1在VSMCs细胞周期的启动.
目的 探討細胞週期蛋白A (Cyclin A)對培養大鼠血管平滑肌細胞(VSMCs)週期起始識彆複閤物1(ORC1)錶達水平的影響.方法 採用組織塊貼片法原代培養的大鼠胸主動脈血管平滑肌細胞,利用雙胸苷使VSMCs達到細胞週期同步化,應用逆轉錄聚閤酶鏈反應技術和流式細胞儀檢測不同細胞週期VSMCs的ORC1和Cyclin A mRNA和蛋白錶達.結果 經鑒定培養的細胞為VSMCs,採用血清饑餓法、雙胸苷阻斷法和鞦水仙素阻抑法分彆穫得瞭處于不同細胞週期的同步化的VSMCs:G0/G1期(89.22 ±3.54)％,G1/S交界期(66.74±7.16)％,S期(63.24±4.06)％,G2/M期(51.64±11.18)％,各期比較差異有統計學意義(P均＜0.01).Cyclin A在靜止狀態對VSMCs的ORC1 mRNA和蛋白無影響,在G2/M期達到高峰[(52.133 3 ±2.122 1)％],與G1/S期[(10.9167±0.531 1)％]、S期[(7.656 7±0.412 4)％]比較,差異有統計學意義(P均＜0.01).ORC1在G2/M期達到最低值[(1.276 7 ±0.161 7)％],與G1/S期[(13.371 0±1.057 3)％]、S期[(3.043 3 ±0.538 0)％]比較,差異有統計學意義(P均＜0.01),可能原因是Cyclin A阻止ORC1結閤在VSMCs染色質上.結論 CyclinA可能調控ORC1在VSMCs細胞週期的啟動.
목적 탐토세포주기단백A (Cyclin A)대배양대서혈관평활기세포(VSMCs)주기기시식별복합물1(ORC1)표체수평적영향.방법 채용조직괴첩편법원대배양적대서흉주동맥혈관평활기세포,이용쌍흉감사VSMCs체도세포주기동보화,응용역전록취합매련반응기술화류식세포의검측불동세포주기VSMCs적ORC1화Cyclin A mRNA화단백표체.결과 경감정배양적세포위VSMCs,채용혈청기아법、쌍흉감조단법화추수선소조억법분별획득료처우불동세포주기적동보화적VSMCs:G0/G1기(89.22 ±3.54)％,G1/S교계기(66.74±7.16)％,S기(63.24±4.06)％,G2/M기(51.64±11.18)％,각기비교차이유통계학의의(P균＜0.01).Cyclin A재정지상태대VSMCs적ORC1 mRNA화단백무영향,재G2/M기체도고봉[(52.133 3 ±2.122 1)％],여G1/S기[(10.9167±0.531 1)％]、S기[(7.656 7±0.412 4)％]비교,차이유통계학의의(P균＜0.01).ORC1재G2/M기체도최저치[(1.276 7 ±0.161 7)％],여G1/S기[(13.371 0±1.057 3)％]、S기[(3.043 3 ±0.538 0)％]비교,차이유통계학의의(P균＜0.01),가능원인시Cyclin A조지ORC1결합재VSMCs염색질상.결론 CyclinA가능조공ORC1재VSMCs세포주기적계동.
Objective To explore the effect of Cyclin A on origin recognition complex-1 (ORC1) expression of rat vascular smooth cells (VSMCs).Methods Primary VSMCs of thoracic aorta in rats was obtained by the adherence method of tissue culture.The synchrony of cell was obtained by the method of double-thymidine block.In different cell cycles of VSMCs,the expression of ORC1 mRNA was determined by RT-PCR and the protein expression of ORC1 was observed by flow cytometry.Results Synchronized VSMCs were obtained and identified by the methods of double-thymidine block,colchicine treatment and serum starvation.Synchronized growth was monitored by flow cytometry.All the synchronized VSMC's distribution ration was (89.22±3.54) ％ at G0/G1 phase,(66.74 ±7.16)％ at G1/S phase,(63.24 ±4.06)％ at S phase and (51.64 ± 11.18)％ at G2/M phase and there was statistically significant difference compared with other phase(P ＜0.01).There was no significant effect of Cyclin A on ORC1 mRNA expression at a quiescent stage of VSMC.At G2/M phase peaked ORC1 was (52.133 3 ± 2.122 1)％,at G1/S phase was(10.916 7 ± 0.531 1)％,at S phase was (7.656 7 ± 0.412 4)％,and there was statistically significant difference (P ＜0.01).At G2/M phase ORC1 downed to the lowest point was (1.276 7 ± 0.161 7) ％,at G1/S phase was (13.371 0 ± 1.057 3)％,at S phase was (3.043 3 ± 0.538 0)％,and there was statistically significant difference (P ＜ 0.01),suggesting that Cyclin A might prevent ORC1 binding the chromatin of VSMCs.Conclusion Cyclin A may be an important regulative factor at the initiation of ORC1 in VSMCs.
