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Inhibition of integrin-linked kinase via a siRNA expression plasmid attenuates connective tissue growth factor-induced human proximal tubular epithelial cells to mesenchymal transition.
Am J Nephrol. 2008; 28(1):143-51.AJ

Abstract

BACKGROUND

Increasing evidence suggests that connective tissue growth factor (CTGF) is involved in the epithelial-to-mesenchymal transition (EMT). The exact intracellular events that drive this process, however, are not fully understood. In this study, we investigated the role of integrin-linked kinase (ILK) in mediating CTGF-induced EMT.

METHODS

The expression of alpha-smooth muscle actin (alpha-SMA) and E-cadherin upon the stimulation by recombinant human CTGF (rhCTGF) in cultured human tubular epithelial cell line (HK-2) was detected by real-time RT-PCR and Western blot. Subsequently, the role of ILK was determined by using ILK siRNA.

RESULTS

rhCTGF increased the mRNA expression of alpha-SMA significantly in a dose- and time-dependent manner, while E-cadherin mRNA decreased in a dose- and time-dependent manner. alpha-SMA protein was up-regulated after stimulation by 5 ng/ml CTGF for 96 h, and increased further after stimulation by 50 ng/ml. An immunocytochemical study showed that alpha-SMA was initially detectable at 48 h, and increased further at 72 h, while there was almost no alpha-SMA immunostaining observed in the control group at the same time point. E-cadherin protein was also down-regulated in a dose-dependent manner. Transfection of HK-2 cells with ILK-siRNA significantly attenuated rhCTGF-induced alpha-SMA induction and E-cadherin repression.

CONCLUSION

Our study suggested that ILK mediated the effect of EMT in proximal tubular epithelial cells stimulated by CTGF.

Authors+Show Affiliations

Institute of Nephrology, Zhong Da Hospital, Southeast University, Nanjing, PR China. liubc64@yahoo.com.cnNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17951996

Citation

Liu, Bi-Cheng, et al. "Inhibition of Integrin-linked Kinase Via a siRNA Expression Plasmid Attenuates Connective Tissue Growth Factor-induced Human Proximal Tubular Epithelial Cells to Mesenchymal Transition." American Journal of Nephrology, vol. 28, no. 1, 2008, pp. 143-51.
Liu BC, Li MX, Zhang JD, et al. Inhibition of integrin-linked kinase via a siRNA expression plasmid attenuates connective tissue growth factor-induced human proximal tubular epithelial cells to mesenchymal transition. Am J Nephrol. 2008;28(1):143-51.
Liu, B. C., Li, M. X., Zhang, J. D., Liu, X. C., Zhang, X. L., & Phillips, A. O. (2008). Inhibition of integrin-linked kinase via a siRNA expression plasmid attenuates connective tissue growth factor-induced human proximal tubular epithelial cells to mesenchymal transition. American Journal of Nephrology, 28(1), 143-51.
Liu BC, et al. Inhibition of Integrin-linked Kinase Via a siRNA Expression Plasmid Attenuates Connective Tissue Growth Factor-induced Human Proximal Tubular Epithelial Cells to Mesenchymal Transition. Am J Nephrol. 2008;28(1):143-51. PubMed PMID: 17951996.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Inhibition of integrin-linked kinase via a siRNA expression plasmid attenuates connective tissue growth factor-induced human proximal tubular epithelial cells to mesenchymal transition. AU - Liu,Bi-Cheng, AU - Li,Min-Xia, AU - Zhang,Jian-Dong, AU - Liu,Xiao-Cong, AU - Zhang,Xiao-Liang, AU - Phillips,Aled O, Y1 - 2007/10/19/ PY - 2007/05/07/received PY - 2007/08/24/accepted PY - 2007/10/24/pubmed PY - 2007/12/14/medline PY - 2007/10/24/entrez SP - 143 EP - 51 JF - American journal of nephrology JO - Am J Nephrol VL - 28 IS - 1 N2 - BACKGROUND: Increasing evidence suggests that connective tissue growth factor (CTGF) is involved in the epithelial-to-mesenchymal transition (EMT). The exact intracellular events that drive this process, however, are not fully understood. In this study, we investigated the role of integrin-linked kinase (ILK) in mediating CTGF-induced EMT. METHODS: The expression of alpha-smooth muscle actin (alpha-SMA) and E-cadherin upon the stimulation by recombinant human CTGF (rhCTGF) in cultured human tubular epithelial cell line (HK-2) was detected by real-time RT-PCR and Western blot. Subsequently, the role of ILK was determined by using ILK siRNA. RESULTS: rhCTGF increased the mRNA expression of alpha-SMA significantly in a dose- and time-dependent manner, while E-cadherin mRNA decreased in a dose- and time-dependent manner. alpha-SMA protein was up-regulated after stimulation by 5 ng/ml CTGF for 96 h, and increased further after stimulation by 50 ng/ml. An immunocytochemical study showed that alpha-SMA was initially detectable at 48 h, and increased further at 72 h, while there was almost no alpha-SMA immunostaining observed in the control group at the same time point. E-cadherin protein was also down-regulated in a dose-dependent manner. Transfection of HK-2 cells with ILK-siRNA significantly attenuated rhCTGF-induced alpha-SMA induction and E-cadherin repression. CONCLUSION: Our study suggested that ILK mediated the effect of EMT in proximal tubular epithelial cells stimulated by CTGF. SN - 1421-9670 UR - https://www.unboundmedicine.com/medline/citation/17951996/Inhibition_of_integrin_linked_kinase_via_a_siRNA_expression_plasmid_attenuates_connective_tissue_growth_factor_induced_human_proximal_tubular_epithelial_cells_to_mesenchymal_transition_ L2 - https://www.karger.com?DOI=10.1159/000110019 DB - PRIME DP - Unbound Medicine ER -