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[Construction and expression of mouse DOC-1R antisense gene vector]
Wei-qiang Zhou1, Li Jiang, Xiu-jie Sheng
1Key Laboratory of Cell Biology, Institute of Medical Molecular Biology, Shenyang 110001, P. R. China. zhouwq@263.net
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.
Method:
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.
Result:
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.
Conclusion:
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.
Insights
The DOC-1R gene, a potential tumor suppressor, was investigated for its effects on normal cell growth. Antisense DOC-1R plasmid construction revealed its ability to inhibit cell replication and colony formation, suggesting therapeutic potential.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Context:
- The DOC-1R gene is recognized as a potential tumor suppressor.
- It interacts with CDK2 and cyclin to regulate the cell cycle.
- Understanding its role in normal cell growth is crucial for cancer research.
Purpose:
- To construct an antisense DOC-1R plasmid.
- To investigate the impact of the DOC-1R gene on normal cell growth and replication.
- To analyze the effects on cell proliferation and colony formation.
Summary:
- Recombinant antisense and sense DOC-1R plasmids were created and transfected into NIH3T3 cells.
- Cell replication and soft agar colony formation assays were performed to assess DOC-1R's function.
- The mouse DOC-1R gene significantly inhibited cell replication and colony formation capacity.
Impact:
- The study demonstrates that the DOC-1R gene acts as a suppressor of normal cell growth.
- Findings provide insights into the mechanisms of normal cell growth and replication.
- This research has implications for tumor treatment and prevention strategies.
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