Abstract
Transforming growth factor-β1 (TGF-β1) is involved in the regulation of trophoblast cell proliferation and invasion. However, the mechanism underlying this process remains unknown, which is predominantly due to the difficulty in obtaining and maintaining primary trophoblast cells in culture over a long period of time. The HTR-8/SVneo cell line is an immortalized trophoblast cell line, which has been reported to exhibit a number of similar characteristics to those of parental trophoblast cells. Therefore, the cell line has been a useful tool for the investigation of placental function and tumor progression. In the present study, the HTR-8/SVneo cell line was used as a model to investigate the TGF-β1/SMAD signaling pathway in the proliferation and invasion of trophoblast cells. The proliferation and invasion ability of HTR-8/SVneo cells was determined using the MTT and Transwell assays, respectively. In addition, reverse transcription polymerase chain reactions were performed to detect the mRNA expression of a panel of known downstream mediators of TGF-β1, including TGF-β receptor I (TβRI), SMAD4, SMAD3, SMAD7 and tissue inhibitor of metalloproteinases-1 (TIMP-1). The results indicated that TGF-β1 promotes the proliferation and invasion of the HTR-8/SVneo cell line at passage 90. Furthermore, the expression of TβRI, SMAD3 and SMAD4 were reduced following treatment with TGF-β1, while the expression of SMAD7 was increased and the expression of TIMP-1 remained unchanged following TGF-β1 treatment. These observations indicated that the effects of TGF-β1 on the proliferation and invasion of the HTR-8/SVneo cell line at passage 90 were different from those of parental trophoblasts, which is in contrast to the results of previous studies. It was concluded that the HTR-8/SVneo cell lines, which have been grown for over 90 passages, do not accurately represent parental trophoblast cells in studies of the TGF-β/SMAD signaling pathway.
Highlights
The proliferation and invasion of the first trimester human trophoblast cell is important in embryonic development
The HTR‐8/SVneo cell line has been established to investigate the biology of normal trophoblast cells as they have been reported to share certain characteristics with their parental cells
The normal trophoblast proliferation and invasiveness of the uterus are strictly regulated processes that are inhibited by Transforming growth factor‐β1 (TGF‐β1) produced locally [6,7,8,9]
Summary
The proliferation and invasion of the first trimester human trophoblast cell is important in embryonic development. Deficient EVT invasion is associated with complications during pregnancy, including intrauterine growth restriction and pre‐eclampsia [1]. The proliferation and invasion of the first trimester human trophoblast cell is influenced by multiple regulatory factors, including growth factors, cytokines, adhesion molecules, proteases, matrix‐derived components and oxygen tension [3,4]. Transforming growth factor‐β1 (TGF‐β1) is involved in trophoblast proliferation and invasion [5,6,7]. TGF‐β receptor I (TβRI) and SMADs are the key downstream mediators of transcriptional responses to TGF‐β. TGF‐β signaling is known to be involved in the regulation of proliferation, differentiation and apoptosis of numerous cells [14]
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