Limits...
Twist-mediated Epithelial-mesenchymal Transition Promotes Breast Tumor Cell Invasion via Inhibition of Hippo Pathway.

Wang Y, Liu J, Ying X, Lin PC, Zhou BP - Sci Rep (2016)

Bottom Line: However, the mechanisms that facilitate the functions of Twist remain unclear.Here we report that Twist overexpression increased expression of PAR1, an upstream regulator of the Hippo pathway; PAR1 promotes invasion, migration, and CSC-like properties in breast cancer by activating the transcriptional co-activator TAZ.Our study indicates that Hippo pathway inhibition is required for the increased migratory and invasiveness ability of breast cancer cells in Twist-mediated EMT.

View Article: PubMed Central - PubMed

Affiliation: Cancer Institute of Integrative Medicine, Zhejiang Academy of Chinese Medicine, Hangzhou, Zhejiang, 310007, China.

ABSTRACT
Twist is a key transcription factor for Epithelial-mesenchymal transition (EMT), which is a cellular de-differentiation program that promotes invasion and metastasis, confers tumor cells with cancer stem cell (CSC)-like characteristics, and increases therapeutic resistance. However, the mechanisms that facilitate the functions of Twist remain unclear. Here we report that Twist overexpression increased expression of PAR1, an upstream regulator of the Hippo pathway; PAR1 promotes invasion, migration, and CSC-like properties in breast cancer by activating the transcriptional co-activator TAZ. Our study indicates that Hippo pathway inhibition is required for the increased migratory and invasiveness ability of breast cancer cells in Twist-mediated EMT.

No MeSH data available.


Related in: MedlinePlus

Overexpression of Twist induces the activation of PAR1 signaling.(A) Graphic representation of the fold change in mRNA levels of Twist, F2R and F2RL2 in Twist-expressing T47D cells compared with control vector cells by real-time PCR. Presented data are the mean ± SD of three separate experiments, with *indicates p < 0.01 when comparing with control values. (B) Western blot analysis for p-TAZ, TAZ and CTGF expression in EMT-induced Twist-expressing T47D cells or T47D cells expressing control vector. Actin served as a loading control. (C) Effect of TAZ siRNA or NTC siRNA on CTGF promoter luciferase activity in Twist-overexpressing T47D cells and T47D cells expressing control vector. Assessments were made after 48 hours in culture. Presented data are mean ± SD of normalized luciferase activities determined from three separate experiments. *indicates p < 0.01 when control siRNA expressed in Twist-T47D cells compared with in vector control cells; and **indicates p < 0.01 when compared expression of TAZ siRNA and control siRNA in Twist-T47D cells. (D) Effect of TAZ siRNA on TAZ and Twist expression in EMT-induced Twist-expressing T47D cells and in T47D cells expressing control vector by western blot analysis. Actin served as a loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4837350&req=5

f5: Overexpression of Twist induces the activation of PAR1 signaling.(A) Graphic representation of the fold change in mRNA levels of Twist, F2R and F2RL2 in Twist-expressing T47D cells compared with control vector cells by real-time PCR. Presented data are the mean ± SD of three separate experiments, with *indicates p < 0.01 when comparing with control values. (B) Western blot analysis for p-TAZ, TAZ and CTGF expression in EMT-induced Twist-expressing T47D cells or T47D cells expressing control vector. Actin served as a loading control. (C) Effect of TAZ siRNA or NTC siRNA on CTGF promoter luciferase activity in Twist-overexpressing T47D cells and T47D cells expressing control vector. Assessments were made after 48 hours in culture. Presented data are mean ± SD of normalized luciferase activities determined from three separate experiments. *indicates p < 0.01 when control siRNA expressed in Twist-T47D cells compared with in vector control cells; and **indicates p < 0.01 when compared expression of TAZ siRNA and control siRNA in Twist-T47D cells. (D) Effect of TAZ siRNA on TAZ and Twist expression in EMT-induced Twist-expressing T47D cells and in T47D cells expressing control vector by western blot analysis. Actin served as a loading control.

Mentions: To understand the molecular mechanisms associated with Twist-induced EMT, we performed cDNA microarray analysis of HMLE and T47D cells that had undergone Twist-mediated EMT (Figs 1A and 2A). The mRNA of two PARs family genes, PAR1 (F2R) and PAR3 (F2RL2), were significantly elevated in both cell lines. These results were confirmed by qRT-PCR (Fig. 5A). Recently, these PAR proteins were identified as upstream regulators of the Hippo pathway, and play a crucial role in breast cancer invasion and metastasis. These data suggest that Twist regulates the Hippo pathway by upregulating PAR expression.


Twist-mediated Epithelial-mesenchymal Transition Promotes Breast Tumor Cell Invasion via Inhibition of Hippo Pathway.

Wang Y, Liu J, Ying X, Lin PC, Zhou BP - Sci Rep (2016)

Overexpression of Twist induces the activation of PAR1 signaling.(A) Graphic representation of the fold change in mRNA levels of Twist, F2R and F2RL2 in Twist-expressing T47D cells compared with control vector cells by real-time PCR. Presented data are the mean ± SD of three separate experiments, with *indicates p < 0.01 when comparing with control values. (B) Western blot analysis for p-TAZ, TAZ and CTGF expression in EMT-induced Twist-expressing T47D cells or T47D cells expressing control vector. Actin served as a loading control. (C) Effect of TAZ siRNA or NTC siRNA on CTGF promoter luciferase activity in Twist-overexpressing T47D cells and T47D cells expressing control vector. Assessments were made after 48 hours in culture. Presented data are mean ± SD of normalized luciferase activities determined from three separate experiments. *indicates p < 0.01 when control siRNA expressed in Twist-T47D cells compared with in vector control cells; and **indicates p < 0.01 when compared expression of TAZ siRNA and control siRNA in Twist-T47D cells. (D) Effect of TAZ siRNA on TAZ and Twist expression in EMT-induced Twist-expressing T47D cells and in T47D cells expressing control vector by western blot analysis. Actin served as a loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837350&req=5

f5: Overexpression of Twist induces the activation of PAR1 signaling.(A) Graphic representation of the fold change in mRNA levels of Twist, F2R and F2RL2 in Twist-expressing T47D cells compared with control vector cells by real-time PCR. Presented data are the mean ± SD of three separate experiments, with *indicates p < 0.01 when comparing with control values. (B) Western blot analysis for p-TAZ, TAZ and CTGF expression in EMT-induced Twist-expressing T47D cells or T47D cells expressing control vector. Actin served as a loading control. (C) Effect of TAZ siRNA or NTC siRNA on CTGF promoter luciferase activity in Twist-overexpressing T47D cells and T47D cells expressing control vector. Assessments were made after 48 hours in culture. Presented data are mean ± SD of normalized luciferase activities determined from three separate experiments. *indicates p < 0.01 when control siRNA expressed in Twist-T47D cells compared with in vector control cells; and **indicates p < 0.01 when compared expression of TAZ siRNA and control siRNA in Twist-T47D cells. (D) Effect of TAZ siRNA on TAZ and Twist expression in EMT-induced Twist-expressing T47D cells and in T47D cells expressing control vector by western blot analysis. Actin served as a loading control.
Mentions: To understand the molecular mechanisms associated with Twist-induced EMT, we performed cDNA microarray analysis of HMLE and T47D cells that had undergone Twist-mediated EMT (Figs 1A and 2A). The mRNA of two PARs family genes, PAR1 (F2R) and PAR3 (F2RL2), were significantly elevated in both cell lines. These results were confirmed by qRT-PCR (Fig. 5A). Recently, these PAR proteins were identified as upstream regulators of the Hippo pathway, and play a crucial role in breast cancer invasion and metastasis. These data suggest that Twist regulates the Hippo pathway by upregulating PAR expression.

Bottom Line: However, the mechanisms that facilitate the functions of Twist remain unclear.Here we report that Twist overexpression increased expression of PAR1, an upstream regulator of the Hippo pathway; PAR1 promotes invasion, migration, and CSC-like properties in breast cancer by activating the transcriptional co-activator TAZ.Our study indicates that Hippo pathway inhibition is required for the increased migratory and invasiveness ability of breast cancer cells in Twist-mediated EMT.

View Article: PubMed Central - PubMed

Affiliation: Cancer Institute of Integrative Medicine, Zhejiang Academy of Chinese Medicine, Hangzhou, Zhejiang, 310007, China.

ABSTRACT
Twist is a key transcription factor for Epithelial-mesenchymal transition (EMT), which is a cellular de-differentiation program that promotes invasion and metastasis, confers tumor cells with cancer stem cell (CSC)-like characteristics, and increases therapeutic resistance. However, the mechanisms that facilitate the functions of Twist remain unclear. Here we report that Twist overexpression increased expression of PAR1, an upstream regulator of the Hippo pathway; PAR1 promotes invasion, migration, and CSC-like properties in breast cancer by activating the transcriptional co-activator TAZ. Our study indicates that Hippo pathway inhibition is required for the increased migratory and invasiveness ability of breast cancer cells in Twist-mediated EMT.

No MeSH data available.


Related in: MedlinePlus