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Crosstalk between the p38 and TGF-β signaling pathways through TβRI, TβRII and Smad3 expression in plancental choriocarcinoma JEG-3 cells.

Tan Y, Xu Q, Li Y, Mao X, Zhang K - Oncol Lett (2014)

Bottom Line: MTT assay was used to detect the effect of TGF-β1 on JEG-3 cell proliferation.The data demonstrated that TGF-β can enhance the viability of JEG-3 cells.Analysis revealed that p38 MAPK is involved in and contributes to the TGF-β pathway, dependent on the regulation of TβRI, TβRII and Smad3.

View Article: PubMed Central - PubMed

Affiliation: Department of Basic Medicine, Chengde Medical College, Chengde, Hebei 067000, P.R. China.

ABSTRACT
Choriocarcinoma is a highly aggressive tumor that develops from germ cells. Some choriocarcinomas originate in the testes or ovaries, while others may develop in the uterus after a normal pregnancy or after miscarriage. The tumor is characterized by early hematogenous spread to distal organs, such as the lung and brain. Transforming growth factor β1 (TGF-β1) is key in regulating tumor cell proliferation and invasion through a variety of Smad-dependent and -independent pathways, including the p38 mitogen-activated protein kinase (MAPK) pathway. There appears to be crosstalk between the TGF-β/Smad and p38 MAPK pathways; however, the molecular mechanisms underlying the crosstalk are not fully understood. The present study validated the role of TGF-β signaling in cancer progression and explored the interaction between Smad and p38 MAPK signaling on transduction mediators in choriocarcinoma using the JEG-3 cell line. MTT assay was used to detect the effect of TGF-β1 on JEG-3 cell proliferation. Cells were treated with p38 MAPK inhibitor and TGF-β receptor inhibitor, followed by TGF-β1, and reverse transcription quantitative real-time polymerase chain reaction was used to examine the transcriptional levels of Smad3 and TGF-β receptors. The data demonstrated that TGF-β can enhance the viability of JEG-3 cells. Blockade of the TGF-β and p38 MAPK pathways attenuated the expression of Smad3, TGF-β receptor type I (TβRI) and TβRII, and inhibited their expression in a dose-dependent manner. Analysis revealed that p38 MAPK is involved in and contributes to the TGF-β pathway, dependent on the regulation of TβRI, TβRII and Smad3. Further investigation of the interactions between the TGF-β and p38 MAPK pathways may offer potential venues for therapeutic intervention for choriocarcinoma.

No MeSH data available.


Related in: MedlinePlus

JEG-3 cells were treated with 5 ng/ml TGF-β1, except the blank and control groups. Cellular proliferation was determined using the MTT assay at 1, 2, 6, 12 and 24 h, respectively (mean ± SD, n=3). TGF-β1, transforming growth factor β1.
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f1-ol-08-03-1307: JEG-3 cells were treated with 5 ng/ml TGF-β1, except the blank and control groups. Cellular proliferation was determined using the MTT assay at 1, 2, 6, 12 and 24 h, respectively (mean ± SD, n=3). TGF-β1, transforming growth factor β1.

Mentions: JEG-3 cells were incubated in six-well plates with an initial concentration of 5×104 cells/ml for 48 h. Wells were divided into six groups as follows: Control, TGF-β1, 1-μM SB203580, 3-μM SB203580, 1-μM LY364947 and 3-μM LY364947 groups. Cells in the control group were cultured with RPMI-1640 only, while the cells in the TGF-β1 groups were treated with 5 ng/ml TGF-β1 and incubate for 2 h. When the cells reached ~80% confluency, they were pretreated with the appropriate concentration of TGF-β1 receptor inhibitor (LY36494; Sigma-Aldrich) and p38 MAPK inhibitor (SB203580; Sigma-Aldrich), and cultured for 2 h. The cells were cultured for 2 h as according to the MTT results (Fig. 1), no obvious effect on proliferation was identified with the treatment of TGF-β1 for 2 h, ensuring that the expression changes of the TGF-β receptors and Smad3 were not affected by changes in cell proliferation levels. Subsequently, 5 ng/ml TGF-β1 (PeproTech, Inc.) was added to each well, with the exception of those in the control group, and incubation was continued for 2 h. Total RNA extraction of JEG-3 cells from each group was performed with TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA). RNA (1 μg) was reverse transcribed using an M-MLV First-Strand cDNA Synthesis kit (Invitrogen Life Technologies) and random oligodeoxynucleotide primers.


Crosstalk between the p38 and TGF-β signaling pathways through TβRI, TβRII and Smad3 expression in plancental choriocarcinoma JEG-3 cells.

Tan Y, Xu Q, Li Y, Mao X, Zhang K - Oncol Lett (2014)

JEG-3 cells were treated with 5 ng/ml TGF-β1, except the blank and control groups. Cellular proliferation was determined using the MTT assay at 1, 2, 6, 12 and 24 h, respectively (mean ± SD, n=3). TGF-β1, transforming growth factor β1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4114612&req=5

f1-ol-08-03-1307: JEG-3 cells were treated with 5 ng/ml TGF-β1, except the blank and control groups. Cellular proliferation was determined using the MTT assay at 1, 2, 6, 12 and 24 h, respectively (mean ± SD, n=3). TGF-β1, transforming growth factor β1.
Mentions: JEG-3 cells were incubated in six-well plates with an initial concentration of 5×104 cells/ml for 48 h. Wells were divided into six groups as follows: Control, TGF-β1, 1-μM SB203580, 3-μM SB203580, 1-μM LY364947 and 3-μM LY364947 groups. Cells in the control group were cultured with RPMI-1640 only, while the cells in the TGF-β1 groups were treated with 5 ng/ml TGF-β1 and incubate for 2 h. When the cells reached ~80% confluency, they were pretreated with the appropriate concentration of TGF-β1 receptor inhibitor (LY36494; Sigma-Aldrich) and p38 MAPK inhibitor (SB203580; Sigma-Aldrich), and cultured for 2 h. The cells were cultured for 2 h as according to the MTT results (Fig. 1), no obvious effect on proliferation was identified with the treatment of TGF-β1 for 2 h, ensuring that the expression changes of the TGF-β receptors and Smad3 were not affected by changes in cell proliferation levels. Subsequently, 5 ng/ml TGF-β1 (PeproTech, Inc.) was added to each well, with the exception of those in the control group, and incubation was continued for 2 h. Total RNA extraction of JEG-3 cells from each group was performed with TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA). RNA (1 μg) was reverse transcribed using an M-MLV First-Strand cDNA Synthesis kit (Invitrogen Life Technologies) and random oligodeoxynucleotide primers.

Bottom Line: MTT assay was used to detect the effect of TGF-β1 on JEG-3 cell proliferation.The data demonstrated that TGF-β can enhance the viability of JEG-3 cells.Analysis revealed that p38 MAPK is involved in and contributes to the TGF-β pathway, dependent on the regulation of TβRI, TβRII and Smad3.

View Article: PubMed Central - PubMed

Affiliation: Department of Basic Medicine, Chengde Medical College, Chengde, Hebei 067000, P.R. China.

ABSTRACT
Choriocarcinoma is a highly aggressive tumor that develops from germ cells. Some choriocarcinomas originate in the testes or ovaries, while others may develop in the uterus after a normal pregnancy or after miscarriage. The tumor is characterized by early hematogenous spread to distal organs, such as the lung and brain. Transforming growth factor β1 (TGF-β1) is key in regulating tumor cell proliferation and invasion through a variety of Smad-dependent and -independent pathways, including the p38 mitogen-activated protein kinase (MAPK) pathway. There appears to be crosstalk between the TGF-β/Smad and p38 MAPK pathways; however, the molecular mechanisms underlying the crosstalk are not fully understood. The present study validated the role of TGF-β signaling in cancer progression and explored the interaction between Smad and p38 MAPK signaling on transduction mediators in choriocarcinoma using the JEG-3 cell line. MTT assay was used to detect the effect of TGF-β1 on JEG-3 cell proliferation. Cells were treated with p38 MAPK inhibitor and TGF-β receptor inhibitor, followed by TGF-β1, and reverse transcription quantitative real-time polymerase chain reaction was used to examine the transcriptional levels of Smad3 and TGF-β receptors. The data demonstrated that TGF-β can enhance the viability of JEG-3 cells. Blockade of the TGF-β and p38 MAPK pathways attenuated the expression of Smad3, TGF-β receptor type I (TβRI) and TβRII, and inhibited their expression in a dose-dependent manner. Analysis revealed that p38 MAPK is involved in and contributes to the TGF-β pathway, dependent on the regulation of TβRI, TβRII and Smad3. Further investigation of the interactions between the TGF-β and p38 MAPK pathways may offer potential venues for therapeutic intervention for choriocarcinoma.

No MeSH data available.


Related in: MedlinePlus