Zhou Wei-qiang, Jiang Li, Sheng Xiu-jie, Zhang Xue
Key Laboratory of Cell Biology, Institute of Medical Molecular Biology, Shenyang 110001, P. R. China.
Ai Zheng. 2002 Mar;21(3):240-4.
BACKGROUND & OBJECTIVE: DOC-1R gene is a candidate tumor suppressor gene that when connected specifically with CDK2 can control the course of cell cycle by restraining the reciprocity of CDK2 and cyclin. The aim of this study was to construct the antisense DOC-1R plasmid and to investigate the effect of DOR-1R gene on the growth of normal cell.
The recombinant antisense plasmid was constructed after screening the expression of DOC-1R gene. Following transfection, the effect of DOC-1R on cell growth was determined by assessing the ability of cell replication and observing soft agar culture.
The growth speed of NIH3T3 transfected by mouse DOC-1R gene was of significant difference from that transfected by empty vector. The pcDNA3-DOC-1R+ vector significantly inhibited the cell replication, while the pcDNA3-DOC-1R- vector stimulated the cell replication. In soft agar culture, the colony formation capacity was decreased in the recombinant sense vector group. The clone formation rate was decreased and the size of the colony formed was smaller as well. In contrast, the colony forming ability was remarkably increased in the antisense vector group. The clone formation rate was increased significantly, compared to that in the sense group.
Mouse DOC-1R gene can significantly inhibit cell growth and colony formation capacity. It will be helpful for the study on the mechanism of normal cell growth and replication as well as for research in tumor treatment and prevention.
DOC-1R基因是一种候选抑癌基因，它与细胞周期蛋白依赖性激酶2（CDK2）特异性结合时，可通过抑制CDK2与细胞周期蛋白的相互作用来控制细胞周期进程。本研究旨在构建反义DOC-1R质粒，并探讨DOC-1R基因对正常细胞生长的影响。
筛选DOC-1R基因表达后构建重组反义质粒。转染后，通过评估细胞复制能力和观察软琼脂培养来确定DOC-1R对细胞生长的影响。
用小鼠DOC-1R基因转染的NIH3T3细胞生长速度与用空载体转染的细胞有显著差异。pcDNA3-DOC-1R+载体显著抑制细胞复制，而pcDNA3-DOC-1R-载体则刺激细胞复制。在软琼脂培养中，重组正义载体组的集落形成能力下降。克隆形成率降低，形成的集落大小也较小。相反，反义载体组的集落形成能力显著增加。与正义组相比，克隆形成率显著提高。
小鼠DOC-1R基因可显著抑制细胞生长和集落形成能力。这将有助于正常细胞生长和复制机制的研究，以及肿瘤治疗和预防的研究。
