[Influences of recombinant adeno-associated virus-mediated Decorin gene transfection on cell cycle and apoptosis of SiHa cells].
OBJECTIVE
Decorin (DCN), a multi-efficiency factor, has many important functions, such as influencing fibril stability, and interacting with extracellular matrix molecules to influence cell adhesion. This study was to investigate influences of DCN on cell cycle and apoptosis of SiHa cells via recombinant adeno-associated virus-mediated DCN gene (rAAV.DCN) transfection.


METHODS
Based on the cDNA sequence of DCN reported by GenBank, DCN segment with open reading frame was cloned into plasmid UF(1) (UF(1)-DCN) after DNA sequencing. rAAV.DCN, containing high titer of DCN, was produced by transfecting into 293 cells with calcium phosphate-DNA sedimentation method, and transfected into SiHa cells, rAAV.LacZ-infected cells and non-infected cells were used as controls. Expression of DCN was determined by Western blot. Cell cycle and apoptosis were analyzed by flow cytometry (FCM).


RESULTS
High titer of rAAV .DCN was produced successfully. After transfected with rAAV.DCN, SiHa cells effectively expressed DCN. The percentage of G(1) phase in rAAV.DCN-transfected cells was significantly higher than that in control cells [(72.3+/-2.3)% vs. (59.5+/-2.5)%, P<0.05], while the percentages of S phase, and G(2) phase in rAAV.DCN-transfected cells were significantly lower than those in control cells [(11.9+/-1.9)% vs. (17.8+/-0.8)%, and (13.4+/-1.1)% vs. (21.5+/-0.9)%, P<0.05]. No change of apoptosis was detected in rAAV.DCN-transfected cells.


CONCLUSIONS
Cell cycle of SiHa cells may be arrested by high expression of DCN.
Zhao
Chun-Xia
CX
Department of Internal Medicine and Gene Therapy Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430-030, P.R. China.
Zhao
Yin-Tao
YT
Wang
Pei-Hua
PH
Xiao
Xiao
X
Wang
Dao-Wen
DW
D017209:Apoptosis; D002453:Cell Cycle; D002460:Cell Line; D045744:Cell Line, Tumor; D058575:Decorin; D000229:Dependovirus / Q000235:genetics*; D004622:Embryo, Mammalian; D016326:Extracellular Matrix Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology; D016366:Open Reading Frames; D010957:Plasmids; D011509:Proteoglycans / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000473:pathology*
C546878:DCN protein, human; D058575:Decorin; D016326:Extracellular Matrix Proteins; D011509:Proteoglycans