中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
12期
2452-2454
,共3页
任泽强%雍红梅%张秀忠%张蓬波%白津%时梅林%郑骏年
任澤彊%雍紅梅%張秀忠%張蓬波%白津%時梅林%鄭駿年
임택강%옹홍매%장수충%장봉파%백진%시매림%정준년
Cul1基因%乳腺癌%增殖%细胞周期
Cul1基因%乳腺癌%增殖%細胞週期
Cul1기인%유선암%증식%세포주기
Cul1 gene%Breast cancer%Proliferation%Cell cycle
目的 探讨下调Cullin1(Cul1)基因对乳腺癌细胞增殖的影响及其机制.方法 利用脂质体将靶向Cul1基因小干扰RNA(siRNA)、对照siRNA分别转染人乳腺癌MDA-MB-231和BT-549细胞,Western blot技术检测Cul1基因表达,细胞计数试剂盒(CCK-8)比色法检测细胞增殖,流式细胞仪检测细胞周期变化,Western blot检测细胞周期蛋白(Cyclin)A、Cyclin D1、Cyclin E以及细胞周期蛋白依赖性激酶抑制因子p21和p27表达.结果 与对照组比较,转染Cull siRNA 48 h后MDA-MB-231和BT-549细胞Cul1蛋白表达明显降低;细胞增殖明显减慢,72 h和96 h更加明显(P＜0.01);Cul1基因干涉6h后2种细胞G1期细胞比例[(29.49±0.75)％、(24.86％±0.52)％]增加,与对照组[(17.39±0.56)％、(12.73％±0.41)％]比较差异有统计学意义(P＜0.01).Cul1基因干涉后2种细胞Cyclin A、Cyclin D1及Cyclin E表达明显减少,而p21和p27表达明显增加,差异均有统计学意义(P＜0.01).结论 Cul1基因干涉后能通过影响细胞周期蛋白及其依赖性激酶抑制因子的表达,使乳腺癌细胞停滞在G1期,最终抑制其增殖.
目的 探討下調Cullin1(Cul1)基因對乳腺癌細胞增殖的影響及其機製.方法 利用脂質體將靶嚮Cul1基因小榦擾RNA(siRNA)、對照siRNA分彆轉染人乳腺癌MDA-MB-231和BT-549細胞,Western blot技術檢測Cul1基因錶達,細胞計數試劑盒(CCK-8)比色法檢測細胞增殖,流式細胞儀檢測細胞週期變化,Western blot檢測細胞週期蛋白(Cyclin)A、Cyclin D1、Cyclin E以及細胞週期蛋白依賴性激酶抑製因子p21和p27錶達.結果 與對照組比較,轉染Cull siRNA 48 h後MDA-MB-231和BT-549細胞Cul1蛋白錶達明顯降低;細胞增殖明顯減慢,72 h和96 h更加明顯(P＜0.01);Cul1基因榦涉6h後2種細胞G1期細胞比例[(29.49±0.75)％、(24.86％±0.52)％]增加,與對照組[(17.39±0.56)％、(12.73％±0.41)％]比較差異有統計學意義(P＜0.01).Cul1基因榦涉後2種細胞Cyclin A、Cyclin D1及Cyclin E錶達明顯減少,而p21和p27錶達明顯增加,差異均有統計學意義(P＜0.01).結論 Cul1基因榦涉後能通過影響細胞週期蛋白及其依賴性激酶抑製因子的錶達,使乳腺癌細胞停滯在G1期,最終抑製其增殖.
목적 탐토하조Cullin1(Cul1)기인대유선암세포증식적영향급기궤제.방법 이용지질체장파향Cul1기인소간우RNA(siRNA)、대조siRNA분별전염인유선암MDA-MB-231화BT-549세포,Western blot기술검측Cul1기인표체,세포계수시제합(CCK-8)비색법검측세포증식,류식세포의검측세포주기변화,Western blot검측세포주기단백(Cyclin)A、Cyclin D1、Cyclin E이급세포주기단백의뢰성격매억제인자p21화p27표체.결과 여대조조비교,전염Cull siRNA 48 h후MDA-MB-231화BT-549세포Cul1단백표체명현강저;세포증식명현감만,72 h화96 h경가명현(P＜0.01);Cul1기인간섭6h후2충세포G1기세포비례[(29.49±0.75)％、(24.86％±0.52)％]증가,여대조조[(17.39±0.56)％、(12.73％±0.41)％]비교차이유통계학의의(P＜0.01).Cul1기인간섭후2충세포Cyclin A、Cyclin D1급Cyclin E표체명현감소,이p21화p27표체명현증가,차이균유통계학의의(P＜0.01).결론 Cul1기인간섭후능통과영향세포주기단백급기의뢰성격매억제인자적표체,사유선암세포정체재G1기,최종억제기증식.
Objective To investigate the role of Cuilinl (Cul1) gene on human breast cancer cell proliferation.Methods The Cull small interfering RNA (siRNA) were transferred into MDA-MB-231 and BT-549 breast cancer cells.The expression of Cul1 gene in both cancer cells were assessed by Western blotting.The proliferation of both cancer cells after Cul1 interference were measured by cell counting kit-8 (CCK-8) assay.The cell cycle of breast cancer cells were detected by flow cytometry.The expression levels of Cyclin family (Cyclin A,Cyclin D1,Cyclin E) and CDK inhibitor proteins (p21 and p27) were detected by Western blotting.Results Cul1 siRNA could silence the expression of Cul1 gene in both breast cancer cells significantly.Cul1 interference could drastically decreased the ability of cell proliferation in both breast cancer cells and rose to its peak level at 96 h.The G1 population in both breast cancer cells treated by Cul1 siRNA [(29.49 ± 0.75) ％,(24.86 ± 0.52) ％] increased significantly than the ceils treated by control siRNA [(17.39 ± 0.56) ％,(12.73 ± 0.41) ％] at 6 h.The expression levels of Cyclin A,Cyclin D1,and Cyclin E proteins were drastically decreased in both breast cancer cells after Cul1 interference,at the same time,p21 and p27 protein expression levels was significantly increased.Conclusion Silence of Cul1 could suppress human breast cancer cell growth through down-regulation of Cyclin A,Cyclin D1,and Cyclin E proteins,up-regulation of p21 and p27 proteins making cell cycle arrest at G1 phase.
