Effect of Silencing SLPI Gene Expression on Differentiation of BeWo Cells
Neelima P. Sidarthan,
Vijayakumar Govindaraj,
Mary Nirmala Sarkar,
A. J. Rao.
Issue:
Volume 1, Issue 1, May 2013
Pages:
1-8
Received:
17 April 2013
Published:
2 May 2013
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.
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 fo...
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Involvement of Pro-Apoptotic and Pro-Autophagic Proteins in Granulosa Cell Death
Escobar M. L.,
Echeverría O .M.,
Casasa A .S.,
García G.,
Aguilar S. J.,
Vázquez-Nin G.H.
Issue:
Volume 1, Issue 1, May 2013
Pages:
9-17
Received:
3 May 2013
Published:
20 May 2013
Abstract: Follicular atresia is a process present in all mammals studied. It involves the oocyte and granulosa cells. The apoptotic cell death has been implicated in follicular atresia. Now it is known that the autophagy is a programmed cell death. In this work, atretic follicles of Wistar rats’ ovaries were analyzed, to evaluate the routes of granulosa cell death during the follicular atresia. The apoptosis and autophagy presence was studied by means of ultrastructural and immunohis-tochemical techniques, and by molecular procedures. During atresia, follicular cells undergo the standard processes of cell death, apoptosis and autophagy, as well as a process in which features of both occur in the same cell. Other processes of cell death affect only granulosa cells and involve such features as contraction of cell volume, an increase of the lumen of the nuclear envelope and the endoplasmic reticulum, and loss of contacts with the oocyte, which is also altered.
Abstract: Follicular atresia is a process present in all mammals studied. It involves the oocyte and granulosa cells. The apoptotic cell death has been implicated in follicular atresia. Now it is known that the autophagy is a programmed cell death. In this work, atretic follicles of Wistar rats’ ovaries were analyzed, to evaluate the routes of granulosa cell...
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