201. Homeobox gene DLX3 regulates forskolin induced trophoblast differentiation
A. Chui A , B. Kalionis A , S. Brennecke A and P. Murthi AUniversity of Melbourne/Royal Women’s Hospital, Department of Obstetrics and Gynaecology, Melbourne, Vic., Australia.
Reproduction, Fertility and Development 20(9) 1-1 https://doi.org/10.1071/SRB08Abs201
Published: 28 August 2008
Abstract
Trophoblast cells carry out important functions required for the development of the normal placenta. Disruption of these functions is associated with significant pregnancy disorders such as fetal growth restriction and pre-eclampsia. Transcription factors regulate trophoblast functions. We are interested in one such class of transcription factors known as homeobox genes. The homeobox gene Distal-less 3 (DLX3) plays a vital role in the development of the mouse placenta (Morasso, Grinberg et al. 1999) and increased levels of DLX3 have been found in placentae affected by human fetal growth restriction (Murthi and Chui, unpubl. data). However, the function of DLX3 in the human placenta is not well established. Here, we investigated whether DLX3 regulates trophoblast differentiation using a plasmid construct to overexpress DLX3 in the human trophoblast cell line, BeWo. Real-time PCR showed a significant increase in DLX3 mRNA (3.1 ± 0.1 v. 1.0 ± 0.2 control, P < 0.05, n = 3), as well as the mRNA of two known markers of differentiation 3-β-hydroxysteroid dehydrogenase (3β-HSD) (8.1 ± 1.8 v. 1.2 ± 0.1 control, P < 0.05, n = 3) and β-human chorionic gonadotropin (β-hCG) (54.9 ± 0.9 v. 49.2 ± 1.6 control, P < 0.05, n = 3). Furthermore, forskolin mediated trophoblast differentiation was verified in BeWo cells. Following forskolin induction, real-time PCR showed a significant increase in the expression of DLX3 mRNA (12.9 ± 1.2 v. 3.8 ± 0.9 control, P < 0.05, n = 4), as well as a significant increase in the mRNA expression of the differentiation markers previously tested, 3β-HSD (28.3 ± 2.4 v. 1.0 ± 0.08 control, P < 0.05, n = 4) and β-hCG (2.3 ± 1.9 v. 30.9 ± 0.08 control, P < 0.001, n = 3). The expression of an additional differentiation marker, syncytin, was also significantly increased (4.0 ± 1.9 v. 1.0 ± 0.08 control, P < 0.05, n = 4). Thus, we have shown that DLX3 is a regulator of human trophoblast cell differentiation, and that forskolin acts through DLX3 to induce trophoblast differentiation.
(1) Morasso, M. I., A. Grinberg et al. (1999).
