Effect of human telomerase reverse transcriptase transfection on differentiation in BeWo choriocarcinoma cells.

Abstract:

:Arrest of proliferation is one of the prerequisites for differentiation of cytotrophoblasts into syncytiotrophoblasts, and thus during differentiation telomerase activity, as well as human telomerase reverse transcriptase (hTERT) expression, is down-regulated. Considering this, it is of interest to investigate whether syncytium formation can be delayed by prolonging the expression of telomerase in cytotrophoblasts. BeWo cells were transfected with pLPC-hTERT retroviral vector and the reverse transcription-polymerase chain reaction analysis for hTERT mRNA concentrations in the transfected cells revealed a several-fold increase in hTERT mRNA compared with the cells transfected with empty vector, and this confirmed that the transfection was successful. An increase in the proliferation, as assessed by bromodeoxyuridine incorporation assay, as well as an increase in mRNA and protein concentration of various cyclins and proliferating cell nuclear antigen, was noticed. The effect of hTERT transfection was also assessed after the addition of forskolin to induce differentiation and it was observed that cell-cell fusion was delayed and differentiation did not occur in hTERT-transfected cells. However, the effects seen were only transient as stable transfection was not possible and the cells were undergoing apoptosis after 72 h, which suggested that apart from hTERT other factors might be important for immortalization of BeWo cells.

journal_name

Reprod Biomed Online

authors

Neelima PS,Rao Rekha M,Rama S,Rao AJ

doi

10.1016/s1472-6483(10)60035-1

subject

Has Abstract

pub_date

2009-06-01 00:00:00

pages

838-49

issue

6

eissn

1472-6483

issn

1472-6491

pii

S1472-6483(10)60035-1

journal_volume

18

pub_type

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