摘要
为了探讨缺乏可测出Ｇ１、Ｇ２期Ｖ７９－８细胞株的细胞周期失调的分子机理，构建了人的周期负调控基因ｐ２１Ｃｉｐ１的可表达重组质粒ｐＸＪＳＣ，转染该细胞系，建立了高表达ｐ２１的Ｖ７９－８－ＳＣ细胞。以３ＨＴｄＲ放射自显影法，流式细胞光度术，细胞增殖曲线测定及免疫印迹技术，检测及比较了与细胞周期变化和增殖有关基因的表达，发现该细胞较对照细胞Ｖ７９－８－ｎｅｏＧ１期明显延长，Ｇ２期也稍有增加。与此同时，Ｇ１期调控的ｃｙｃｌｉｎＤ１、ＣＤＫ４、Ｉｄ基因表达明显下降，Ｇ２期调控基因ｃｙｃｌｉｎＢ１的表达也有所下降。结果表明：此细胞系的细胞周期缺陷可能由于ｐ２１Ｃｉｐ１。
To explore the molecular mechanism of cell cycle control disorder of the cell line, V 79-8 which has no detectable G1 and G2 phases, we constructed human negative cell cycle control gene p21Cip1, eukaryoticaly expressing recombinant pXJSC, and transfected it into V 79-8 to establish V 79-8 sc cell line, which had higher p21 expression. By using 3HTdR autoradiography, FCM, cell growth curve measurement, and Northern botl techniques to measure and compare the changes of cell cycle time,and cell proliferation related gene expression, we found that V 79-8 sc cells have obviously lengthened G1 phase and slightly lengthened G2 phase comparing with control V 79-8 neo cells. At the same time, the expression of G1 regulators, such as cyclin D1, CDK4, and Id are obviously decreased and the G2 regulator cyclinB1 is also decreased. From all the results, we recognized that the failure in cell cycle might be due to the discordance of p21cip1, CDKs/cyclins and its related gene expression.
出处
《解剖学报》
CAS
CSCD
北大核心
1998年第3期264-269,I001,共7页
Acta Anatomica Sinica
基金
国家自然科学基金
