中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
5期
351-353
,共3页
代军%张培海%刘培淑%曲红华
代軍%張培海%劉培淑%麯紅華
대군%장배해%류배숙%곡홍화
卵巢肿瘤%细胞周期蛋白D%逆转录聚合酶链反应%流式细胞术%细胞增殖
卵巢腫瘤%細胞週期蛋白D%逆轉錄聚閤酶鏈反應%流式細胞術%細胞增殖
란소종류%세포주기단백D%역전록취합매련반응%류식세포술%세포증식
Ovarian neoplasms%Cyclin D%RT-PCR%Flow cytometry%Cell proliferation
目的 探讨细胞周期素D1( cyclinD1)在上皮性卵巢癌3AO细胞中的表达,分析其表达与3AO细胞生长、增殖的关系.方法 将人正常卵巢上皮细胞及卵巢癌3AO细胞进行体外培养,用逆转录酶多聚酶链式反应(RT-PCR)及流式细胞术(FCM)方法检测受不同浓度顺铂作用前后,细胞中cyclinD1基因及蛋白的表达情况,并观察细胞凋亡及细胞周期的变化.结果 人卵巢上皮细胞中未检测到cyclinD1基因及蛋白的表达,而3AO细胞中存在cyclinD1基因的异常扩增及过表达；受顺铂作用后,3AO细胞琼脂糖凝胶电泳显示cyclinD1基因的mRNA显带由明变暗,逐渐变模糊,并且显带变暗程度与顺铂细胞作用浓度呈正比.流式细胞术直标单抗分析,3AO细胞中cyclinD1蛋白呈阳性过表达；受顺铂作用前后,cyclinD1蛋白表达水平不同,平均值分别是105.9、15.42、8.59;细胞凋亡率不同,分别是0.63％,9.08％,27.41％；细胞周期发生了改变,细胞增殖指数分别是38.83％,44.54％,37.31％,差异有统计学意义(P＜0.01).结论 卵巢癌3AO细胞中存在cyclinD1基因的异常扩增及过表达,cyclinD1基因及蛋白的阳性表达下降程度与顺铂细胞作用浓度呈正比；cyclinD1的表达与卵巢癌细胞的生长及增殖活性密切相关.
目的 探討細胞週期素D1( cyclinD1)在上皮性卵巢癌3AO細胞中的錶達,分析其錶達與3AO細胞生長、增殖的關繫.方法 將人正常卵巢上皮細胞及卵巢癌3AO細胞進行體外培養,用逆轉錄酶多聚酶鏈式反應(RT-PCR)及流式細胞術(FCM)方法檢測受不同濃度順鉑作用前後,細胞中cyclinD1基因及蛋白的錶達情況,併觀察細胞凋亡及細胞週期的變化.結果 人卵巢上皮細胞中未檢測到cyclinD1基因及蛋白的錶達,而3AO細胞中存在cyclinD1基因的異常擴增及過錶達；受順鉑作用後,3AO細胞瓊脂糖凝膠電泳顯示cyclinD1基因的mRNA顯帶由明變暗,逐漸變模糊,併且顯帶變暗程度與順鉑細胞作用濃度呈正比.流式細胞術直標單抗分析,3AO細胞中cyclinD1蛋白呈暘性過錶達；受順鉑作用前後,cyclinD1蛋白錶達水平不同,平均值分彆是105.9、15.42、8.59;細胞凋亡率不同,分彆是0.63％,9.08％,27.41％；細胞週期髮生瞭改變,細胞增殖指數分彆是38.83％,44.54％,37.31％,差異有統計學意義(P＜0.01).結論 卵巢癌3AO細胞中存在cyclinD1基因的異常擴增及過錶達,cyclinD1基因及蛋白的暘性錶達下降程度與順鉑細胞作用濃度呈正比；cyclinD1的錶達與卵巢癌細胞的生長及增殖活性密切相關.
목적 탐토세포주기소D1( cyclinD1)재상피성란소암3AO세포중적표체,분석기표체여3AO세포생장、증식적관계.방법 장인정상란소상피세포급란소암3AO세포진행체외배양,용역전록매다취매련식반응(RT-PCR)급류식세포술(FCM)방법검측수불동농도순박작용전후,세포중cyclinD1기인급단백적표체정황,병관찰세포조망급세포주기적변화.결과 인란소상피세포중미검측도cyclinD1기인급단백적표체,이3AO세포중존재cyclinD1기인적이상확증급과표체；수순박작용후,3AO세포경지당응효전영현시cyclinD1기인적mRNA현대유명변암,축점변모호,병차현대변암정도여순박세포작용농도정정비.류식세포술직표단항분석,3AO세포중cyclinD1단백정양성과표체；수순박작용전후,cyclinD1단백표체수평불동,평균치분별시105.9、15.42、8.59;세포조망솔불동,분별시0.63％,9.08％,27.41％；세포주기발생료개변,세포증식지수분별시38.83％,44.54％,37.31％,차이유통계학의의(P＜0.01).결론 란소암3AO세포중존재cyclinD1기인적이상확증급과표체,cyclinD1기인급단백적양성표체하강정도여순박세포작용농도정정비；cyclinD1적표체여란소암세포적생장급증식활성밀절상관.
Objective To explore.the expression of cyclinD1 in ovarian carcinoma cell 3AO and analyze its relationship with the proliferation of ovarian cancer cell.Methods Human ovarian epithelium and ovarian cancer cells 3AO were cultured in vito.CyclinD1 genes and proteins were detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry before and after the activation of cell 3AO by cis-platinum. Also the cell activity and cell cycle were observed. Results Abnormal gene amplification and over-expressions of cyclinD1 were found in ovarian cancer cell while the expression of cyclinD1 was negative in normal ovarian epithelium.Under cis-platinum,different expressions of cyclinD1 genes were found in 3AO by RT-PCR.The higher the concentrations of cisplatin,the lower expressions of cyclinD1 genes.By flow cytometry,it was also found that there were lower expressions of cyclinD1 protein in 3AO under cisplatin than without it. With the rising concentrations of cisplatin,the low expressions of cyclinD1 protein in 3AO were detected.The mean numbers were 105.9,15.42 and 8.59,the cell apoptotic rates 0.63％,9.08％ and 27.41％ and the proliferation index (PI) numbers 38.83％,44.54％,37.31％.The differences were statistically significant (P ＜ 0.01 ).Conclusion The up-regulation of cyclinD1 is detected in ovarian cancer cell.A positive correlation is found between the lower expressions of cyclinD1 and the concentration of cicplatin.There is a close relationship between the expressions of cyclinD1 and ovarian cancer cell activity.
