[Over-expression of HIF-1alpha induces EMT of human prostate cancer cells].
OBJECTIVE
To determine whether human prostate cancer cell lines undergo epithelial-mesenchymal transition (EMT) and become more invasive when induced by HIF-1alpha, and to explore the underlying molecular mechanism.


METHODS
The cell line LNCaP, appropriate for the HIF-1alpha induction test, was screened out from 4 different EMT-negative prostate cell lines through vimentin gene detection by RT-PCR. The recombinant plasmid pCDNA3. 1(-)/HIF-1alpha was constructed and transfected into LNCaP with the Lipofectamine 2000 system. The control plasmid pCDNA3.1 (-) was transfected by the same method. The positive clone cells were selected by G418 and confirmed by Western blot and immunofluorescence staining. Then a Transwell polycarbonate filter, coated with 100 micol Matrigel at 1:20 dilution in the serum-free medium, was used to analyze the invasive potency. The expression of E-cadherin and vimentin was detected by Western blot.


RESULTS
Among the 4 different EMT-negative cell lines, LNCaP was the only one that expressed the vimentin gene but not protein. The expression of HIF1alpha was obviously higher in LNCaP/HIF1alpha than in LNCaP/pCDNA3. 1 (- an LNCaP. The number of the LNCaP/HIF1alpha cells that penetrated through the Transwell polycarbonate filter was significantly larger than that of the LNCaP and LNCaP/pCDNA3. 1(-) cells. Compared with the LNCaP/pCDNA3.1(-) and LNCaP cells, the expression of vimentin was up-regulated, while that of E-cadherin down-regulated, in LNCaP/HIF1alpha.


CONCLUSIONS
The over-expression of HIF-1alpha could induce EMT in the human prostate carcinoma cell line LNCaP and enhance its invasiveness through E-cadherin and vimentin regulation.
Luo
Yong
Y
Department of Urology, Beijing Anzhen Hospital Affiliated to Capital Medical University, Beijing 100029, China. luoyongxjtu@sohu.com
He
Da-Lin
DL
Jiang
Yong-Guang
YG
Li
Ming-Chuan
MC
Ning
Liang
L
Shen
Shu-Lin
SL
D015820:Cadherins / Q000096:biosynthesis; D045744:Cell Line, Tumor / Q000378:metabolism / Q000473:pathology; D004848:Epithelium / Q000473:pathology*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D008297:Male; D009361:Neoplasm Invasiveness; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014746:Vimentin / Q000096:biosynthesis
D015820:Cadherins; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D014746:Vimentin