Ding, J. X., Feng, Y. J., Yao, L. Q., Yu, M., Jin, H. Y., Yin, L. H. (2006) The reinforcement of invasion in epithelial ovarian cancer cells by 17?-Estradiol is associated with up-regulation of Snail. Gynecologic Oncology, 103 (2). pp. 623-630. ISSN 00908258
Abstract
Objective.: The transcription factor Snail, which is implicated in the triggering of epithelial-mesenchymal transitions (EMT), plays an important role in adhesion, invasion and metastasis of tumor cells. In the present study, we assessed 17?-Estradiol (E2)'s effect on Snail, E-cadherin and MMP-2 expression of epithelial ovarian cancer cell line ES-2 and SKOV3. Then we induced Snail gene silencing by RNA interference to explore the effect of E2 on E-cadherin and MMP-2 expression when Snail gene expression was blocked. Methods.: Treated by 10-8 M E2, Snail, E-cadherin and MMP-2 mRNA expression of the cells was measured by RT-PCR; Snail, MMP-2 protein expression was detected by IHC; and MMP-2 activity was determined by Zymography. E-cadherin protein level was measured by Western blot. We constructed the small interfering dsRNA expression vector (pRNAT-U6.1/Neo-Snail) targeting Snail gene, as well as a negative control vector (pRNAT-U6.1/Neo-Neg). Then the cells were transiently transfected with the vectors. Western blot and zymography were conducted to determine E-cadherin protein level and matrix metalloproteinase activity of the cells transfected with pRNAT-U6.1/Neo-Snail or pRNAT-U6.1/Neo-Neg after treated with E2 for 24 h. Results.: The expression of ER? mRNA and protein was negative in ES-2 cells and positive in SKOV3 cells, and ER? expression was positive in both cell lines. 10-8 mol/l E2 elevated expression of Snail and MMP-2 mRNA and protein in both ES-2 and SKOV3 cells, and reduced expression of E-cadherin mRNA and protein in SKOV3 cells. While in the RNAi group transfected with the small interfering dsRNA expression vector (pRNAT-U6.1/Neo-Snail) targeting Snail gene, E2 treatment did not have a significant effect on MMP-2 activity or E-cadherin protein in ES-2 and SKOV3 cells. Conclusions.: 17?-Estradiol increased Snail expression in both ER?-negative ES-2 cells and ER?-positive SKOV3 cells independent of the existence of ER?. The increase of MMP-2 expression in ES-2 and SKOV3 cells and decrease of E-cadherin expression in SKOV3 cells induced by E2 were associated with up-regulation of Snail. © 2006 Elsevier Inc. All rights reserved.
| Item Type: | Paper |
|---|---|
| Subjects: | diseases & disorders > cancer bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene silencing bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > transcription factor |
| CSHL Authors: | |
| Communities: | CSHL labs > McCombie lab |
| Depositing User: | CSHL Librarian |
| Date: | 2006 |
| Date Deposited: | 15 Dec 2011 16:48 |
| Last Modified: | 13 Apr 2018 15:21 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/22784 |
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