[Enhanced expression of human vimentin intermediate filaments in hepatocellular carcinoma cells decreases their proliferative and invasive abilities in vitro].
OBJECTIVE
Expression of vimentin in carcinoma cells is a marker for poor prognosis in patients. The aim of this investigation was to assess the influence of vimentin on the characteristics of carcinoma cells.


METHODS
The full-length vimentin gene open reading frame (1401 base pairs) was cloned into the plasmid vector pcDNA 3.1 (+), and these vectors were used to stably transfect the human hepatocellular carcinoma HepG2 cell line. Vimentin gene expression was evaluated by RT-PCR and Western blot. Proliferative activity and invasive potential of tumor cells were determined by the CellTiter 96 aqueous one solution cell proliferation assay and BioCoat GFR Matrigel invasion chamber, respectively.


RESULTS
DNA sequencing and restriction endonuclease digestion analysis demonstrated that the recombinant vector was correctly cloned. The stable cell line demonstrated a higher vimentin RNA and protein expression. However, both proliferative and invasive abilities of the cells were reduced in vitro ( P < 0.05).


CONCLUSIONS
A recombinant plasmid pcDNA3. 1-VIM is successfully constructed and a carcinoma cell line HepG2-pV highly expressing vimentin is obtained. Recombinant vimentin protein suppresses the proliferative and invasive abilities of HepG2 cells, suggesting that it might decrease malignant phenotype of tumor cells in vitro. This work makes a foundation for further study on the relationship between vimentin and biological phenotype of carcinoma cells.
Li
Zu-mao
ZM
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Wen
Yan-jun
YJ
Yang
Hong-bin
HB
Qin
Gang
G
Tian
Ling
L
Deng
Hong-xin
HX
Wen
Bin
B
D002465:Cell Movement; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006801:Humans; D009361:Neoplasm Invasiveness; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014746:Vimentin / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger; D011994:Recombinant Proteins; D014746:Vimentin