Influence of connective tissue growth factor antisense oligonucleotide on angiotensin II-induced epithelial mesenchymal transition in HK2 cells.
OBJECTIVE
The present study was designed to further investigate the effect of connective tissue growth factor antisense oligonucleotide (CTGF-AS) on angiotensin II (Ang II)-induced tubular cell epithelial mesenchymal transition (EMT) in vitro.


METHODS
The human proximal tubular cell line (HK2) was grown in Dulbecco's modified Eagle's medium containing 10% heat inactivated fetal calf serum. After being rested in serum-free medium for 24 h, the influence of CTGF-AS (20 mug/mL) on Ang II-induced (0.1 micromol/L) CTGF mRNA and the protein expression were examined by using reverse transcription-polymerase chain reaction and indirect-immunofluorescence. The effect of CTGF-AS on Ang II-induced cellular ultrastructure was observed using a transmissive electronic microscope. The expression of alpha-smooth action (alpha-SMA) was assayed by immunocytochemistry. In all experiments, the control group was treated with scrambled oligonucleotide.


RESULTS
It was shown that Ang II significantly induced the increasing expression of CTGF mRNA and protein (P<0.01, respectively), which were significantly abolished by treatment with CTGF-AS. After stimulating cells with Ang II, the cellular ultrastructure showed mesenchymal features. These effects were partially inhibited by CTGF-AS. Ang II significantly resulted in the expression of alpha-SMA in time dependent manner, which was markedly attenuated by the treatment with CTGF-AS (P<0.01, respectively). In contrast, no similar effects were observed in the control group treated with scrambled oligonucleotide.


CONCLUSIONS
Ang II-induced EMT in human proximal tubular epithelial cells (PTC) can be attenuated by treatment with CTGF-AS. Our data provides further evidence that CTGF might be involved in Ang II-induced EMT in PTC.
Chen
Long
L
Institute of Nephrology, Zhong Da Hospital, Southeast University School of Medicine, Nanjing 210009, China.
Liu
Bi-Cheng
BC
Zhang
Xiao-Liang
XL
Zhang
Jian-Dong
JD
Liu
Hong
H
Li
Min-Xia
MX
D000199:Actins / Q000378:metabolism; D000804:Angiotensin II / Q000008:administration & dosage / Q000494:pharmacology*; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000096:biosynthesis* / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000235:genetics; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis
D000199:Actins; D000804:Angiotensin II; D055513:Connective Tissue Growth Factor; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C526317:CCN2 protein, human; C526316:CCN2 protein, rat; C498862:Acta2 protein, rat