Effects of STI571 and p27 gene clone on proliferation and apoptosis of K562 cells.
OBJECTIVE
To investigate the combined effect of STI571 and p27 gene clone on the regulation of proliferation, cell cycle and apoptosis of K562 cell line.


METHODS
p27 gene was obtained by RT-PCR, and its sequence was approved to be correct. Then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line. p27-pcDNA3.1-K562 cell clone was screened by G418 after transfection, p27 protein was identified by Western blot. MTT was used to detect the survival rate of the cell. Flow cytometry was used to detect cell cycle and apoptosis index.


RESULTS
The expression of p27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. A strong inhibition of cell proliferation was observed in p27-pcDNA3.1-K562 cells as compared with that of the control (pcDNA3.1-K562 cells). The cells at G0/G1 phase were significantly increased, and cells at S phase were greatly declined. The apoptosis index was increased greatly after p27-pcDNA3.1-K562 cells were treated with STI571, and survival rate of the cell was markedly declined (0.35-0.58, P<0.05-0.048 vs STI571-K562 cell, 0.35-0.72, P<0.01-0.001 vs p27-K562 cell).


CONCLUSIONS
p27 and STI571 have a synergistic action on inhibition of proliferation and induction of apoptosis on K562 cells.
Wang
Wei
W
Department of Hematology, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
Yao
Li-Bo
LB
Liu
Xin-Ping
XP
Feng
Qi
Q
Shang
Zhen-Chuan
ZC
Cao
Yun-Xin
YX
Sun
Bing-Zhong
BZ
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D001549:Benzamides; D018797:Cell Cycle Proteins / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D050760:Cyclin-Dependent Kinase Inhibitor p27; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D000068877:Imatinib Mesylate; D066298:In Vitro Techniques; D020014:K562 Cells; D004915:Leukemia, Erythroblastic, Acute / Q000188:drug therapy*; D008969:Molecular Sequence Data; D010879:Piperazines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics*
D000970:Antineoplastic Agents; D001549:Benzamides; D018797:Cell Cycle Proteins; D010879:Piperazines; D011743:Pyrimidines; D025521:Tumor Suppressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D000068877:Imatinib Mesylate