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.
| Published in | Cell Biology (Volume 1, Issue 1) |
| DOI | 10.11648/j.cb.20130101.11 |
| Page(s) | 1-8 |
| Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
| Copyright |
Copyright © The Author(s), 2013. Published by Science Publishing Group |
SLPI, siRNA, Cell Differentiation, Cytotrophoblast, Syncytiotrophoblast
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APA Style
Neelima P. Sidarthan, Vijayakumar Govindaraj, Mary Nirmala Sarkar, A. J. Rao. (2013). Effect of Silencing SLPI Gene Expression on Differentiation of BeWo Cells. Cell Biology, 1(1), 1-8. https://doi.org/10.11648/j.cb.20130101.11
ACS Style
Neelima P. Sidarthan; Vijayakumar Govindaraj; Mary Nirmala Sarkar; A. J. Rao. Effect of Silencing SLPI Gene Expression on Differentiation of BeWo Cells. Cell Biol. 2013, 1(1), 1-8. doi: 10.11648/j.cb.20130101.11
AMA Style
Neelima P. Sidarthan, Vijayakumar Govindaraj, Mary Nirmala Sarkar, A. J. Rao. Effect of Silencing SLPI Gene Expression on Differentiation of BeWo Cells. Cell Biol. 2013;1(1):1-8. doi: 10.11648/j.cb.20130101.11
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Download@article{10.11648/j.cb.20130101.11,
author = {Neelima P. Sidarthan and Vijayakumar Govindaraj and Mary Nirmala Sarkar and A. J. Rao.},
title = {Effect of Silencing SLPI Gene Expression on Differentiation of BeWo Cells},
journal = {Cell Biology},
volume = {1},
number = {1},
pages = {1-8},
doi = {10.11648/j.cb.20130101.11},
url = {https://doi.org/10.11648/j.cb.20130101.11},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.cb.20130101.11},
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.},
year = {2013}
}
TY - JOUR T1 - Effect of Silencing SLPI Gene Expression on Differentiation of BeWo Cells AU - Neelima P. Sidarthan AU - Vijayakumar Govindaraj AU - Mary Nirmala Sarkar AU - A. J. Rao. Y1 - 2013/05/02 PY - 2013 N1 - https://doi.org/10.11648/j.cb.20130101.11 DO - 10.11648/j.cb.20130101.11 T2 - Cell Biology JF - Cell Biology JO - Cell Biology SP - 1 EP - 8 PB - Science Publishing Group SN - 2330-0183 UR - https://doi.org/10.11648/j.cb.20130101.11 AB - 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. VL - 1 IS - 1 ER -
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