Effect of adenovirus-mediated p27 gene expression on the proliferation and apoptosis of HL-60 and Raji cell lines.
OBJECTIVE
To explore the possible effect of p27 gene expression on the proliferation and apoptosis of HL-60 and Raji cell lines.


METHODS
The infections of HL-60 and Raji cells were performed by the adenovirus-mediated p27 gene transfection approach. The efficiency of Adp27 infection and the expression of p27 mRNA and protein were evaluated by X-gal staining, RT-PCR and flow cytometry. The proliferation and apoptosis of HL-60 and Raji cells were estimated by using trypan blue staining, MTT assay, Annexin V/PI and DNA ladder electrophoresis.


RESULTS
The infection efficiency of HL-60 and Raji cells were 40.3 and 32%, respectively; RT-PCR and flow cytometry showed that there were significant expressions of p27 mRNA and protein of HL-60 and Raji cells infected by Adp27, while HL-60 cells themselves showed only faint p27 mRNA and protein, and Raji cells hardly presented p27 mRNA and protein. The strong proliferation inhibitions, which were in a time-dependent manner for HL-60 and Raji cells infected by Adp27, were indicated by cell growth curve and MTT assay. After 72 h infection of HL-60 and Raji cells by Adp27, the Annexin V+/PI- apoptotic cell rates were 46.9 and 35.7% respectively, which were significantly increased compared with control group (4.7 and 5.6% respectively). The typical DNA ladder bands were detectable in HL-60 and Raji cells after 48 h of Adp27 infection.


CONCLUSIONS
The infection of HL-60 and Raji cells by means of adenoviral vector-mediated p27 gene could evidently inhibit cellular proliferation and promote cell apoptosis, which would provide experimental evidence for gene therapy of leukemia/lymphoma using adenovirus-mediated p27 gene approach.
Wang
Qinhong
Q
Department of Hematology, HuaShan Hospital, Medical Center of FuDan University, FuDan University, Shanghai, China. qinhong_wang@yahoo.com.cn
Zhang
Min
M
Fang
Huahua
H
Nie
Xiaoxuan
X
Gao
Li
L
Liu
Yan
Y
Xie
Yanghui
Y
Xie
Yi
Y
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000096:biosynthesis / Q000235:genetics*; D002455:Cell Division / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004247:DNA / Q000032:analysis; D004587:Electrophoresis, Agar Gel / Q000379:methods; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D018922:HL-60 Cells; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics*
D018797:Cell Cycle Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004247:DNA