新基因Mip1的过表达对去血清培养条件下C_2C_(12)细胞生长周期的影响

Effects of New Gene Mip1 Over-expression on the Cell Growth Cycle of C_2C_(12) Cells Cultured Under Condition of Serum Withdrawal

【目的】建立稳定表达新基因Mip1的细胞株 ,并探讨正常和应激状态下Mip1过表达对细胞生长周期的影响。【方法】脂质体转染Mip1的真核表达质粒于C2 C12 细胞中 ,G4 18筛选阳性克隆 ,Westernblot鉴定高表达克隆 ,流式细胞术检测正常和去血清状态下Mip1转基因细胞株和空载体转基因细胞株的生长周期分布情况。【结果】建立了稳定高表达新基因Mip1的C2 C12 细胞株 ,并发现Mip1的过表达可减轻去血清对C2 C12 细胞的生长抑制作用。【结论】Mip1在应激状态下可能具有促细胞生长的功能。

Objective To establish the cell strain capable of stably expressing new gene Mip1 and explore the effect of Mip1 over-expression on cell growth cycle during normal and stress state.The eukaryotic expression plasmids of Mip1 were transfected into C 2C 12 cells using lipofectin and positive clones were screened by G418. The cell clones expressing Mip1 in high level were identified with the Western blots. The growth cycle distribution of Mip1 transgenic strain and empty vector transgenic strain under normal and stress condition was detected by flow cytometry.C 2C 12 cells strain capable of stably expressing new gene Mip1 in high level was established. The overexpression of Mip1 reduced the inhibitory effect of serum withdrawal on the growth of C 2C 12 cells.[Conclusion]Mip1 may stimulate the proliferation and growth of C 2C 12 cells under stress conditions.