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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、ＣＤＫｓ／ｃｙｃｌｉｎｓ及其相关基因的表达失调所致
In order to investigate the molecular mechanism of cell cycle dysfunction in V79-8 cell line G1 and G2, we constructed an expressible recombinant plasmid pXJSC of human cycle negative regulation gene p21Cip1 and transfected this cell line to establish a highly expressed p21 V79-8-SC cells. The expression of genes related to cell cycle and proliferation was detected and compared by 3HTdR autoradiography, flow cytometry, cell proliferation curve and Western blotting. The results showed that the cells were significantly more than the control V79-8-neoG1 Prolongation, G2 phase also increased slightly. Meanwhile, the expression of cyclinD1, CDK4 and Id genes in G1 phase decreased significantly, and the expression of cyclinB1 gene in G2 phase also decreased. The results showed that the cell cycle defects in this cell line may be due to the imbalanced expression of p21Cip1, CDKs / cyclins and their related genes