TY - JOUR
T1 - CDK1 inhibition facilitates formation of syncytiotrophoblasts and expression of human Chorionic Gonadotropin
AU - Ullah, Rahim
AU - Dar, Saira
AU - Ahmad, Tanvir
AU - de Renty, Christelle
AU - Usman, Mohammad
AU - DePamphilis, Melvin L.
AU - Faisal, Amir
AU - Shahzad-ul-Hussan, Syed
AU - Ullah, Zakir
N1 - KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: The project was primarily supported by the faculty initiative fund by Lahore University of Management Sciences and part of the work done at NIH was supported by intramural research program of NIH.
PY - 2018/5/17
Y1 - 2018/5/17
N2 - Aims
The human placental syncytiotrophoblast (STB) cells play essential roles in embryo implantation and nutrient exchange between the mother and the fetus. STBs are polyploid which are formed by fusion of diploid cytotrophoblast (CTB) cells. Abnormality in STBs formation can result in pregnancy-related disorders. While a number of genes have been associated with CTB fusion the initial events that trigger cell fusion are not well understood. Primary objective of this study was to enhance our understanding about the molecular mechanism of placental cell fusion.
Methods
FACS and microscopic analysis was used to optimize Forskolin-induced fusion of BeWo cells (surrogate of CTBs) and subsequently, changes in the expression of different cell cycle regulator genes were analyzed through Western blotting and qPCR. Immunohistochemistry was performed on the first trimester placental tissue sections to validate the results in the context of placental tissue. Effect of Cyclin Dependent Kinase 1 (CDK1) inhibitor, RO3306, on BeWo cell fusion was studied by microscopy and FACS, and by monitoring the expression of human Chorionic Gonadotropin (hCG) by Western blotting and qPCR.
Results
The data showed that the placental cell fusion was associated with down regulation of CDK1 and its associated cyclin B, and significant decrease in DNA replication. Moreover, inhibition of CDK1 by an exogenous inhibitor induced placental cell fusion and expression of hCG.
Conclusion
Here, we report that the placental cell fusion can be induced by inhibiting CDK1. This study has a high therapeutic significance to manage pregnancy related abnormalities.
AB - Aims
The human placental syncytiotrophoblast (STB) cells play essential roles in embryo implantation and nutrient exchange between the mother and the fetus. STBs are polyploid which are formed by fusion of diploid cytotrophoblast (CTB) cells. Abnormality in STBs formation can result in pregnancy-related disorders. While a number of genes have been associated with CTB fusion the initial events that trigger cell fusion are not well understood. Primary objective of this study was to enhance our understanding about the molecular mechanism of placental cell fusion.
Methods
FACS and microscopic analysis was used to optimize Forskolin-induced fusion of BeWo cells (surrogate of CTBs) and subsequently, changes in the expression of different cell cycle regulator genes were analyzed through Western blotting and qPCR. Immunohistochemistry was performed on the first trimester placental tissue sections to validate the results in the context of placental tissue. Effect of Cyclin Dependent Kinase 1 (CDK1) inhibitor, RO3306, on BeWo cell fusion was studied by microscopy and FACS, and by monitoring the expression of human Chorionic Gonadotropin (hCG) by Western blotting and qPCR.
Results
The data showed that the placental cell fusion was associated with down regulation of CDK1 and its associated cyclin B, and significant decrease in DNA replication. Moreover, inhibition of CDK1 by an exogenous inhibitor induced placental cell fusion and expression of hCG.
Conclusion
Here, we report that the placental cell fusion can be induced by inhibiting CDK1. This study has a high therapeutic significance to manage pregnancy related abnormalities.
UR - http://hdl.handle.net/10754/627926
UR - https://www.sciencedirect.com/science/article/pii/S0143400418302285
UR - http://www.scopus.com/inward/record.url?scp=85047058496&partnerID=8YFLogxK
U2 - 10.1016/j.placenta.2018.05.003
DO - 10.1016/j.placenta.2018.05.003
M3 - Article
SN - 0143-4004
VL - 66
SP - 57
EP - 64
JO - Placenta
JF - Placenta
ER -