TY - JOUR
T1 - Effect of human telomerase reverse transcriptase transfection on differentiation in BeWo choriocarcinoma cells
AU - Neelima, P. S.
AU - Rao Rekha, M.
AU - Rama, S.
AU - Rao, A. J.
N1 - Funding Information:
One of the authors, AJ Rao, is thankful to the Department of Science and Technology (Government of India) for the Rajaramanna fellowship (DST). The authors also wish to thank the Indian Institute of Science, Bangalore, the Mellon Foundation, USA, and CONRAD, USA, for the financial assistance provided during the course of the work, and Mrs Shailaja and Mrs Nirmala Singh, respectively, for typing and critically reviewing the manuscript.
PY - 2009/6
Y1 - 2009/6
N2 - 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.
AB - 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.
KW - BeWo
KW - Proliferation
KW - Transfection
KW - hTERT
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U2 - 10.1016/S1472-6483(10)60035-1
DO - 10.1016/S1472-6483(10)60035-1
M3 - Article
C2 - 19490790
AN - SCOPUS:68249116774
SN - 1472-6483
VL - 18
SP - 838
EP - 849
JO - Reproductive BioMedicine Online
JF - Reproductive BioMedicine Online
IS - 6
M1 - 3691
ER -