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Alpha or beta human chorionic gonadotropin knockdown decrease BeWo cell fusion by down-regulating PKA and CREB activation
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DOI：10.1038/srep11210.png)
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Abstract
En 中文
The aim of the present study is to delineate the role of human chorionic gonadotropin (hCG) in trophoblast fusion. In this direction, using shRNA lentiviral particles, alpha- nd beta-hCG silenced 'BeWo' cell lines were generated. Treatment of both alpha-and beta-hCG silenced BeWo cells with either forskolin or exogenous hCG showed a significant reduction in cell fusion as compared with control shRNA treated cells. Studies by qRT-PCR, Western blotting and immunofluorescence revealed down-regulation of fusion-associated proteins such as syncytin-1 and syndecan-1 in the alpha- nd beta-hCG silenced cells. Delineation of downstream signaling pathways revealed that phosphorylation of PKA and CREB were compromised in the silenced cells whereas, no significant changes in p38MAPK and ERK1/2 phosphorylation were observed. Moreover, beta-catenin activation was unaffected by either alpha- or beta-hCG silencing. Further, inhibition of PKA by H89 inhibitor led to a significant decrease in BeWo cell fusion but had no effect on beta-catenin activation suggesting the absence of non-canonical beta-catenin stabilization via PKA. Interestingly, canonical activation of beta-catenin was associated with the up-regulation of Wnt 10b expression. In summary, this study establishes the significance of hCG in the fusion of trophoblastic BeWo cells, but there may be additional factors involved in this process.
Keywords:
TROPHOBLAST DIFFERENTIATION
PLACENTAL-LACTOGEN
EXPRESSION
SYNCYTIN
CATENIN
PHOSPHORYLATION
STIMULATION
SYNDECAN-1
RECEPTORS
CD98
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