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Tight junction protein 1 is regulated by transforming growth factor-β and contributes to cell motility in NSCLC cells.

Lee SH, Paek AR, Yoon K, Kim SH, Lee SY, You HJ - BMB Rep (2015)

Bottom Line: Tight junction protein 1 (TJP1), a component of tight junction, has been reported to play a role in protein networks as an adaptor protein, and TJP1 expression is altered during tumor development.SB431542, a type-I TGF-β receptor inhibitor, as well as SB203580, a p38 kinase inhibitor, significantly abrogated the effect of TGF-β on TJP1 expression.When TJP1 expression was reduced by shRNA lentiviral particles in A549 cells (A549-sh TJP1), wound healing was much lower than in cells infected with control viral particles.

View Article: PubMed Central - PubMed

Affiliation: Cancer Cell and Molecular Biology Branch, Div. of Cancer Biology, National Cancer Center, Goyang 410-769; Division of Molecular Life Sciences, Ewha Womans University, Seoul 120-750, Korea.

ABSTRACT
Tight junction protein 1 (TJP1), a component of tight junction, has been reported to play a role in protein networks as an adaptor protein, and TJP1 expression is altered during tumor development. Here, we found that TJP1 expression was increased at the RNA and protein levels in TGF-β-stimulated lung cancer cells, A549. SB431542, a type-I TGF-β receptor inhibitor, as well as SB203580, a p38 kinase inhibitor, significantly abrogated the effect of TGF-β on TJP1 expression. Diphenyleneiodonium, an NADPH oxidase inhibitor, also attenuated TJP1 expression in response to TGF-β in lung cancer cells. When TJP1 expression was reduced by shRNA lentiviral particles in A549 cells (A549-sh TJP1), wound healing was much lower than in cells infected with control viral particles. Taken together, these data suggest that TGF-β enhances TJP1 expression, which may play a role beyond structural support in tight junctions during cancer development.

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TGF-β induces TJP1 expression in A549 lung carcinoma cells. Subconfluent cells were exposed to TGF-β for 24 h, harvested, lysed, and analyzed for TJP1, E-cadherin, N-cadherin, and GAPDH expression by (A) RT-PCR and (B) real-time PCR in A549 cells. Subconfluent cells were exposed to (C) various concentrations of TGF-β for 24 h or to (D) 50 pM TGF-β for various time periods, as indicated, in A549 cells, and analyzed for TJP1, E-cadherin, N-cadherin, and β-actin expression by immunoblotting. All results are representative of at least three independent experiments. Data are expressed as means ± SE. Statistical significance was assessed using paired Student’s t-tests (*P < 0.0001).
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Figure 001: TGF-β induces TJP1 expression in A549 lung carcinoma cells. Subconfluent cells were exposed to TGF-β for 24 h, harvested, lysed, and analyzed for TJP1, E-cadherin, N-cadherin, and GAPDH expression by (A) RT-PCR and (B) real-time PCR in A549 cells. Subconfluent cells were exposed to (C) various concentrations of TGF-β for 24 h or to (D) 50 pM TGF-β for various time periods, as indicated, in A549 cells, and analyzed for TJP1, E-cadherin, N-cadherin, and β-actin expression by immunoblotting. All results are representative of at least three independent experiments. Data are expressed as means ± SE. Statistical significance was assessed using paired Student’s t-tests (*P < 0.0001).

Mentions: Previously, we showed that TGF-β has an effect on EMT and motility in lung cancer cells (22). To verify the effect of TGF-β on cell adhesion and junctional proteins in A549 cells, we first examined the expression of E-cadherin and N-cadherin in TGF-β-stimulated A549 lung cancer cells (Fig. 1A, B). TGF-β increased N-cadherin expression and decreased E-cadherin expression in A549 lung cancer cells, as expected. In terms of tight junction proteins, TJP1 was increased in response to TGF-β (Fig. 1A), while TJP3 was decreased at the RNA level (data not shown). Because TJP1 decreased in response to TGF-β in various cancer cell lines (15, 23, 24), we conducted additional experiments to confirm the effects of TGF-β on TJP1 expression in A549 lung cancer cells. As shown in Fig. 1C and D, TGF-β induced TJP1 expression, beginning at 10 pM and peaking at 50 pM TGF-β. In time-course experiments, TJP1 expression reached a maximum at 24 h in A549 lung cancer cells. Induction of TJP1 in response to TGF-β was also seen in NCI-H596 and A427 human lung carcinoma cell lines (Fig. 4C), suggesting a role for TGF-β on TJP1 expression in lung cancer.


Tight junction protein 1 is regulated by transforming growth factor-β and contributes to cell motility in NSCLC cells.

Lee SH, Paek AR, Yoon K, Kim SH, Lee SY, You HJ - BMB Rep (2015)

TGF-β induces TJP1 expression in A549 lung carcinoma cells. Subconfluent cells were exposed to TGF-β for 24 h, harvested, lysed, and analyzed for TJP1, E-cadherin, N-cadherin, and GAPDH expression by (A) RT-PCR and (B) real-time PCR in A549 cells. Subconfluent cells were exposed to (C) various concentrations of TGF-β for 24 h or to (D) 50 pM TGF-β for various time periods, as indicated, in A549 cells, and analyzed for TJP1, E-cadherin, N-cadherin, and β-actin expression by immunoblotting. All results are representative of at least three independent experiments. Data are expressed as means ± SE. Statistical significance was assessed using paired Student’s t-tests (*P < 0.0001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4352613&req=5

Figure 001: TGF-β induces TJP1 expression in A549 lung carcinoma cells. Subconfluent cells were exposed to TGF-β for 24 h, harvested, lysed, and analyzed for TJP1, E-cadherin, N-cadherin, and GAPDH expression by (A) RT-PCR and (B) real-time PCR in A549 cells. Subconfluent cells were exposed to (C) various concentrations of TGF-β for 24 h or to (D) 50 pM TGF-β for various time periods, as indicated, in A549 cells, and analyzed for TJP1, E-cadherin, N-cadherin, and β-actin expression by immunoblotting. All results are representative of at least three independent experiments. Data are expressed as means ± SE. Statistical significance was assessed using paired Student’s t-tests (*P < 0.0001).
Mentions: Previously, we showed that TGF-β has an effect on EMT and motility in lung cancer cells (22). To verify the effect of TGF-β on cell adhesion and junctional proteins in A549 cells, we first examined the expression of E-cadherin and N-cadherin in TGF-β-stimulated A549 lung cancer cells (Fig. 1A, B). TGF-β increased N-cadherin expression and decreased E-cadherin expression in A549 lung cancer cells, as expected. In terms of tight junction proteins, TJP1 was increased in response to TGF-β (Fig. 1A), while TJP3 was decreased at the RNA level (data not shown). Because TJP1 decreased in response to TGF-β in various cancer cell lines (15, 23, 24), we conducted additional experiments to confirm the effects of TGF-β on TJP1 expression in A549 lung cancer cells. As shown in Fig. 1C and D, TGF-β induced TJP1 expression, beginning at 10 pM and peaking at 50 pM TGF-β. In time-course experiments, TJP1 expression reached a maximum at 24 h in A549 lung cancer cells. Induction of TJP1 in response to TGF-β was also seen in NCI-H596 and A427 human lung carcinoma cell lines (Fig. 4C), suggesting a role for TGF-β on TJP1 expression in lung cancer.

Bottom Line: Tight junction protein 1 (TJP1), a component of tight junction, has been reported to play a role in protein networks as an adaptor protein, and TJP1 expression is altered during tumor development.SB431542, a type-I TGF-β receptor inhibitor, as well as SB203580, a p38 kinase inhibitor, significantly abrogated the effect of TGF-β on TJP1 expression.When TJP1 expression was reduced by shRNA lentiviral particles in A549 cells (A549-sh TJP1), wound healing was much lower than in cells infected with control viral particles.

View Article: PubMed Central - PubMed

Affiliation: Cancer Cell and Molecular Biology Branch, Div. of Cancer Biology, National Cancer Center, Goyang 410-769; Division of Molecular Life Sciences, Ewha Womans University, Seoul 120-750, Korea.

ABSTRACT
Tight junction protein 1 (TJP1), a component of tight junction, has been reported to play a role in protein networks as an adaptor protein, and TJP1 expression is altered during tumor development. Here, we found that TJP1 expression was increased at the RNA and protein levels in TGF-β-stimulated lung cancer cells, A549. SB431542, a type-I TGF-β receptor inhibitor, as well as SB203580, a p38 kinase inhibitor, significantly abrogated the effect of TGF-β on TJP1 expression. Diphenyleneiodonium, an NADPH oxidase inhibitor, also attenuated TJP1 expression in response to TGF-β in lung cancer cells. When TJP1 expression was reduced by shRNA lentiviral particles in A549 cells (A549-sh TJP1), wound healing was much lower than in cells infected with control viral particles. Taken together, these data suggest that TGF-β enhances TJP1 expression, which may play a role beyond structural support in tight junctions during cancer development.

Show MeSH
Related in: MedlinePlus