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. 2012 Dec;4(6):1344-1348.
doi: 10.3892/ol.2012.906. Epub 2012 Sep 11.

The impact of TGF-β1 on the mRNA expression of TβR I, TβR II, Smad4 and the invasiveness of the JEG-3 placental choriocarcinoma cell line

Yuhong Li  1 , Qian Xu, Zhuo Zhang, Shaochen Liu, Changhua Shi, Yusi Tan
Affiliations

The impact of TGF-β1 on the mRNA expression of TβR I, TβR II, Smad4 and the invasiveness of the JEG-3 placental choriocarcinoma cell line

Yuhong Li et al. Oncol Lett. 2012 Dec.

Abstract

Human choriocarcinoma is one of the most aggressive malignant tumors characterized by early hematogenous spread to lung and brain tissues, and may be a cause of death in patients. Choriocarcinoma may occur following pregnancy and during implantation; however, trophoblastic invasion in human pregnancy is tightly regulated. The transforming growth factor-beta 1 (TGF-β1) has been suggested to play a role in controlling this process. In this study, we investigated the impact of TGF-β1 on invasion, as well as its sites of action in the TGF-β1/Smad pathway using a JEG-3 choriocarcinoma cell line. Following the treatment of cells with different doses of TGF-β1, cell invasion was observed. We also detected the expression of TGF-β receptor type I (TβR I) and TGF-β receptor type II (TβR II), Smad4, matrix metalloprotease (MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)-1 in JEG-3 cells. Our data demonstrated that TGF-β1 promoted the invasive capability of JEG-3 cells depending on the downregulation of TβR I, TβR II, Smad4 and the upregulation of MMP-9 and TIMP-1. These observations suggest that TGF-β1 may play a critical role in the initiation of the trophoblastic invasion process.

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Figures

Figure 1.
Figure 1.
The JEG-3 cells were treated with TGF-β1 for 48 h and their invasive capacity was subsequently detected using the Transwell invasion assay. The total number of transmigrated cells was obtained by counting the number of dyed cells with a light microscope. Exogenous TGF-β1 specifically promoted the invasive capacity of JEG-3 cells. The data are presented as the means ± SD (n=5; *P<0.05 when compared with TGF-β1 0 μg/l; #P<0.05 when compared with any other group except 0 μg/l).
Figure 2.
Figure 2.
Regulatory effects of different doses of TGF-β1 on TβR I, TβR II and Smad4 mRNA expression levels in JEG-3 cells. (A) RT-PCR analysis of TβR I, TβR II and Smad4 mRNA levels in JEG-3 cells cultured in different concentrations of TGF-β1; (B,C,D) Statistical analyses. *P<0.05 when compared with TGF-β1 0 μg/l; #P<0.05 when compared with any other group except 0 μg/l.
Figure 3.
Figure 3.
TGF-β1 upregulated MMP-9 and TIMP-1 protein expressions in JEG-3 with increasing of TGF-β1 concentrations. (A) Western blot assay of MMP-9 and TIMP-1 protein expressions in different concentrations of TGF-β1 in the JEG-3 cells. (B,C) MMP-9 and TIMP-1 protein expression. *P<0.05 when compared with TGF-β1 0 μg/l; #P<0.05 when compared with TGF-β1 100 μg/l.
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