[Construction and expression of mouse DOC-1R antisense gene vector].
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Zhou
Wei-qiang
WQ
Key Laboratory of Cell Biology, Institute of Medical Molecular Biology, Shenyang 110001, P. R. China. zhouwq@263.net
Jiang
Li
L
Sheng
Xiu-jie
XJ
Zhang
Xue
X
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D016373:DNA, Antisense / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D016147:Genes, Tumor Suppressor / Q000502:physiology; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000235:genetics* / Q000502:physiology; D014162:Transfection
D016373:DNA, Antisense; C485718:Doc-1r protein, mouse; D015513:Oncogene Proteins