Effect of silencing SLPI gene expression on differentiation of BeWo cells

Sidhartan, Neelima P., Govindaraj, Vijayakumar, Sarkar, Mary Nirmala and Rao, A. J. (2013) Effect of silencing SLPI gene expression on differentiation of BeWo cells. Cell Biology International, . doi:10.11648/j.cb.20130101.11


Author Sidhartan, Neelima P.
Govindaraj, Vijayakumar
Sarkar, Mary Nirmala
Rao, A. J.
Title Effect of silencing SLPI gene expression on differentiation of BeWo cells
Journal name Cell Biology International   Check publisher's open access policy
ISSN 1065-6995
1095-8355
Publication date 2013-05-02
Year available 2013
Sub-type Article (original research)
DOI 10.11648/j.cb.20130101.11
Open Access Status
Total pages 8
Place of publication Oxford, United Kingdom
Publisher John Wiley & Sons
Collection year 2014
Language eng
Abstract In our previous studies which involved genomic profiling by DD-RTPCR and microarray during forskolin induced differentiation of uninucleated cytotrophoblasts into multinucleated syncytiotrophoblasts using BeWo chorionic carcinoma cells as a model, the expression of one of the transcripts namely, Secretory Leucocyte Protease Inhibitor (SLPI) was found to be very high (10-15 fold) in syncytiotrophoblasts compared to the expression in cytotrophoblasts. SLPI is a protein of 12kDa molecular weight and a variety of activities which include protease inhibition, anti inflammatory and anti microbial activity have been attributed to it. In view of this, an attempt was made to investigate the role of SLPI during differentiation of cytotrophoblasts in to syncytiotrophoblasts. The expression of SLPI in BeWo choriocarcinoma cells was inhibited by use of specific oligos designed. Based on the preliminary study two oligos the use of which resulted in maximum inhibition of expression of SLPI more than 75% by 72hrs as assessed by RT-PCR and Western blot were employed in this study. Inhibition of SLPI expression by siRNA resulted in inhibition of morphological differentiation of BeWo cells. This was also reflected functionally by increase in the protease activity as assessed by gelatin zymography. The observation that the expression of two differentiation markers namely Endoglin and hCG also decreased following silencing suggest a role for SLPI in differentiation of cytotrophoblasts to syncytiotrophoblast. RT-PCR analysis for the proliferation markers Cyclin A2 and PCNA in the SLPI silenced cells revealed an increase in their expression. In contrast analysis for differentiation markers GADD45A, DNA-PK,ADRP, and MAP Kinase revealed a decrease in their expression. These results suggest an important role for SLPI during differentiation of cytotrophoblasts into syncytiotrophoblasts and are of significance in that silencing of a single gene can disrupt this differentiation process and establish the importance of SLPI during differentiation process per se.
Keyword SLPI
siRNA
Cell Differentiation
Cytotrophoblast
Syncytiotrophoblast
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online: 2 May 2013.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
School of Biomedical Sciences Publications
 
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Created: Mon, 27 May 2013, 14:39:24 EST by Neelima Pottekkat Sidharthan on behalf of School of Biomedical Sciences