[Inducing of epithelial mesenchymal transition of HK-2 cells by connective tissue growth factor in vitro].
OBJECTIVE
To investigate the role of connective tissue growth factor (CTGF) in epithelial mesenchymal transition of HK-2 cells in vitro.


METHODS
HK-2 cells were randomly divided into two groups: (1) control group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum only; and (2) experimental group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum and recombinant CTGF at a final concentration of 5 microg/L. The cells were collected at 72 h time points. Direct immunofluorescence staining and immunohistochemistry were used to evaluate the E-cadherin, Vimentin, alpha-SMA and ERK2 in cells. Western-blotting was used to detect the E-cadherin, Vimentin and ERK2 protein expression. Boyden Chamber was used to detect the migration of tubular endothelium at 1 d, 3 d and 5 d.


RESULTS
There were less E-cadherin but more Vimentin expressed in cells of the experimental group. The presence of alpha-SMA was detected at 48 h with peak at 72 h in the cells of the experimental group. On the first day, the cellular migration in the two groups showed no difference. However, after 3 days, the transformed cells migrated surpassed the control group with peak at the 5th day [(45.0+/-1.1):(14.0+/-1.2), P<0.05)].


CONCLUSIONS
Connective tissue growth factor induces mesenchymal transformation of HK-2 cells, in which the ERK2 signaling pathway may play an important role.
Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhao
Z
Ding
Zhi-shui
ZS
Chen
Xi-lin
XL
Chen
Hui
H
Guo
Nian-qiao
NQ
Gong
D000199:Actins / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D055513:Connective Tissue Growth Factor / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D008648:Mesoderm / Q000166:cytology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D011897:Random Allocation; D015398:Signal Transduction; D014746:Vimentin / Q000378:metabolism
C541116:ACTA2 protein, human; D000199:Actins; D015820:Cadherins; D014746:Vimentin; D055513:Connective Tissue Growth Factor; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1