[Inhibition of proliferation of human vascular smooth muscle cells by overexpression of p21 gene].
OBJECTIVE
To observe the apoptosis of human vascular smooth muscle cells (HVSMCs) transfected with p21 gene and investigate its potential mechanism.


METHODS
An adenoviral expression vector with full-length cDNA of p21 gene insert (Ad-p21) was constructed and transfected into HVSMC. A mock vector, pAdeno-X-lac Z constructed with the same method, was transfected into HVSMCs as control. Western blotting and DS-PAGE were used to detect the expression of p21 gene. Fluorescence staining was used to observe the changes in nuclei of apoptotic cells and calculate the cell survival rate. Expression of tissue transglutaminase (tTG) in HVSMC was examined by DIG-marked probe.


RESULTS
Fluorescence staining showed marked fragmentation of nucleus and pyknosis of chromatin in pAdeno-X-p21 transfected cells. The expression of tTG was at a higher level in pAdeno-X-p21 transfected cells than in normal cells and pAdeno-X-lac transfected HVSMCs.


CONCLUSIONS
Overexpression of p21 gene inhibits the proliferation of HVSMC through apoptosis induced by p21 which may relate with overexpression of tTG.
Department of Surgery, Huashan Hospital, Fudan University, Shanghai 200040, China.
Xueli
X
Zhang
Qunhua
Q
Zhang
Ping
P
Li
Bo
B
Yu
Tieping
T
Wang
Yunsong
Y
Zhu
Yanling
Y
Zhang
Duan
D
Cai
D017209:Apoptosis / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D014162:Transfection; D011503:Transglutaminases / Q000235:genetics / Q000378:metabolism*
C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D012333:RNA, Messenger; D011503:Transglutaminases