[Arresting effect of p16 and dll4 transfection on cell cycle of K562 cells].
This study was purposed to investigate the expression and role of eukaryotic expression vector containing p16, dll4 genes in leukemia K562 cells. A vector pBudCE4.1-16-dll4 containing wild type p16cDNA and dll4cDNA was designed and constructed, then this vector was transfected into leukemia K562 cells by using lipofectamine 2000. The expression of p16 and dll4 genes was detected by Western blot, the cell growth curve and cell cycle were determined by CCK-8 kit and flow cytometry respectively. The results showed that the recombinant plasmid pBudCE4.1-16-dll4 was constructed and transfected into K562 cells in vitro successfully. The expression of exogenous P16 and Dll4 proteins could be detected in K562 cells. After transfection for 48 hours, the K562 cells were arrested in G(1) phase, the cell count increased in G(0)/G(1) phase and reduced in S phase, the cell proliferation decreased as compared with control. It is concluded that the p16 and dll4 genes can simultaneously express in K562 cells transfected with recombinant plasmid pBudCE4.1-16-dll4 in vitro which results in G(0)/G(1) arrest and reduces cell proliferation.
Shen
Jie-Fang
JF
Department of Rheumatology and Hematology, The First Hospital Affiliated to Fujian Medical University, Fuzhou 350004, Fujian Province, China.
Rui
Hong-Bing
HB
Su
Jin-Zi
JZ
Kang
Ri-Hui
RH
Lin
Jun-Fang
JF
D017209:Apoptosis; D002453:Cell Cycle; D049109:Cell Proliferation; D019942:Genes, p16; D005822:Genetic Vectors; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics*; D020014:K562 Cells; D007938:Leukemia / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D010957:Plasmids; D014162:Transfection
D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C064045:delta protein