[Inhibitory effect of p16, p53 transfection on leukemic cell lines K562 and HL-60].
This study was purposed to construct a vector containing human suppressor gene p53 and p16, and to investigate their expression and effect on K562 and HL-60 cells. pBudCE4.1-53-16 is a vector designed for simultaneous expression of human suppressor gene p53 and p16 in mammalian cell line. After transfection into K562 cells with lipofectamine(TM) 2000, the expression of p53 and p16 genes was detected by Western blot and immunocytochemical method. The growth curve, apoptosis, cell cycle were assayed by CCK-8 and flow cytometry. The results showed that the recombinant plasmid pBudCE4.1-53-16 was constructed successfully and were verified by PCR and restriction analysis. The expression of P53 and P16 protein could be detected after transfection into leukemia cells (K562 and HL-60) for 48 hours. As compared with control group, the cell proliferation in experimental group was inhibited, the cells were arrested in G0 phase and apoptotic cells increased (p<0.001). It is concluded that the recombinant plasmid pBudCE4.1-53-16 has been established. p16 and p53 in the recombinant plasmid pBudCE4.1-53-16 synchronously express in leukemic cells after transfection in vitro for 2 days and results in reduced proliferation, G0 arrest and apoptosis increase.
Chen
Qi
Q
Department of Hematology, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, Fujian Province, China.
Shen
Jie-Fang
JF
Rui
Hong-Bing
HB
Su
Jin-Zi
JZ
Zhuo
Guang-Sheng
GS
Kang
Ri-Hui
RH
Lin
Jun-Fang
JF
D017209:Apoptosis / Q000235:genetics; D002453:Cell Cycle / Q000235:genetics*; D049109:Cell Proliferation; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D015870:Gene Expression; D016158:Genes, p53; D005822:Genetic Vectors; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D010957:Plasmids; D014162:Transfection
D019941:Cyclin-Dependent Kinase Inhibitor p16