[Suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA].
OBJECTIVE
To study the suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA.


METHODS
In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA (pAP) were transfected into Hep-2 cells by using cationic liposome (lipofectamine 2000) and divided into three groups: non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid, pAP group. Semi quantify RT-PCR, western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor (bFGF).


RESULTS
It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph beta Apr-neol transfected group ( P < 0.01). In the presence of 1 microg/L of bFGF in low serum (0.1% FCS), the pAP transfected cells showed a reduced proliferation rate (MTT assay) while the wild type cells and ph beta Apr-neol transfected cells grew rapidly.


CONCLUSIONS
The growth of Hep-2 cells could be inhibited significantly by perlecan anti-sense cDNA plasmids transfection.
Chen
Guangli
G
Department of Otolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Gong
Shusheng
S
Ding
Juan
J
Liu
Yingpeng
Y
Luo
Linghui
L
Wang
Jianting
J
Chen
Pei
P
Yan
Kaisheng
K
D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018076:DNA, Complementary; D005822:Genetic Vectors; D019812:Heparan Sulfate Proteoglycans / Q000235:genetics*; D006801:Humans; D007822:Laryngeal Neoplasms / Q000235:genetics / Q000473:pathology; D010957:Plasmids; D014162:Transfection
D018076:DNA, Complementary; D019812:Heparan Sulfate Proteoglycans; C071927:perlecan