[Inhibition effect of antisense Bmi-1 on Jurkat cells].
OBJECTIVE
To investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.


METHODS
The antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.


RESULTS
The growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.


CONCLUSIONS
Antisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.
Dalian Medical University, China.
Wei-hong
WH
Liu
Xiu-xiang
XX
Meng
Dan-dan
DD
Liu
Lu-juan
LJ
Shan
Xin-yu
XY
Zhao
D002453:Cell Cycle; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D006801:Humans; D019169:Jurkat Cells; D009687:Nuclear Proteins / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D063150:Polycomb Repressive Complex 1; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics*; D014162:Transfection
C069077:BMI1 protein, human; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D063150:Polycomb Repressive Complex 1