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Overexpression of ETV4 protein in triple-negative breast cancer is associated with a higher risk of distant metastasis.

Yuan ZY, Dai T, Wang SS, Peng RJ, Li XH, Qin T, Song LB, Wang X - Onco Targets Ther (2014)

Bottom Line: ETV4 messenger ribonucleic acid was more than five-fold upregulated in TNBC tissue compared with the control tissue.ETV4 overexpression was found in 57.0% of 135 TNBC cases.Overexpression of ETV4 protein was associated with an advanced stage and a higher proportion of positive lymph node and lymphovascular invasion.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncology in South China, Guangzhou, People's Republic of China ; Collaborative Innovation Center for Cancer Medicine, Guangzhou, People's Republic of China ; Department of Medical Oncology, Sun Yat-Sen University Cancer Center, Guangzhou, People's Republic of China.

ABSTRACT

Background: Patients with triple-negative breast cancer (TNBC) present a higher probability of distant metastasis and lack of effective targeted therapy. ETS translocation variant 4 (ETV4) is an ETS (E-26) transcription factor and has been associated with tumor metastasis. However, the clinical and functional significance of ETV4 in TNBC still remains unclear.

Methods: A human tumor metastasis polymerase chain reaction array was used to profile differential expression of tumor metastasis-related genes in TNBC tissue. Real-time reverse transcription and Western blot analyses were performed to verify ETV4 expression in TNBC cells and tissue. Immunohistochemistry was used to detect expression of ETV4 protein in 135 TNBC tissue samples for association between ETV4 protein expression and clinical outcomes.

Results: A total total of eight upregulated (CCL7, KISS1, MET, MMP7, NR4A3, ETV4, TIMP3, and TSHR) and three downregulated (ITGA7, SSTR, and MMP2) genes were identified between TNBC tissue and the luminal subtype of breast cancer tissue. ETV4 messenger ribonucleic acid was more than five-fold upregulated in TNBC tissue compared with the control tissue. ETV4 overexpression was found in 57.0% of 135 TNBC cases. Overexpression of ETV4 protein was associated with an advanced stage and a higher proportion of positive lymph node and lymphovascular invasion. Patients with an ETV4-overexpressed tumor had a significantly higher risk of developing distant metastasis (P<0.0001) and shorter overall survival and disease-free survival. Overexpression of ETV4 protein was an independent predictor of short disease-free survival of TNBC patients (P=0.021).

Conclusion: Overexpression of ETV4 protein increases risk of developing distant metastasis and results in a poor prognosis for TNBC patients. Thus, ETV4 might be a novel target for developing an alternative therapeutic strategy for prevention of TNBC distant metastasis.

No MeSH data available.


Related in: MedlinePlus

Verification of ETS translocation variant 4 (ETV4) overexpression in triple-negative breast cancer (TNBC) cell lines and tissue.Notes: (A) Western blot analysis of ETV4 protein expression in TNBC cell lines and hormone receptor-positive cell lines. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B) Real-time reverse transcription analysis of ETV4 messenger ribonucleic acid (mRNA) expression in TNBC cell lines and hormone receptor-positive cell lines. Transcript levels were normalized to GAPDH. (C) Western blot analysis of ETV4 protein expression in TNBC tissue (T) and adjacent normal breast tissue (ANT); GAPDH was used as a loading control. (D) Real-time reverse transcription analysis of ETV4 mRNA expression in TNBC tissue and adjacent normal breast tissue; expression data were normalized to GAPDH. Error bars represent the mean ± standard deviation from three independent experiments; *P<0.05. (E) Immunohistochemistry staining of corresponding TNBC tissue (T) and adjacent normal breast tissue (ANT).Abbreviation: ER, estrogen receptor.
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f2-ott-7-1733: Verification of ETS translocation variant 4 (ETV4) overexpression in triple-negative breast cancer (TNBC) cell lines and tissue.Notes: (A) Western blot analysis of ETV4 protein expression in TNBC cell lines and hormone receptor-positive cell lines. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B) Real-time reverse transcription analysis of ETV4 messenger ribonucleic acid (mRNA) expression in TNBC cell lines and hormone receptor-positive cell lines. Transcript levels were normalized to GAPDH. (C) Western blot analysis of ETV4 protein expression in TNBC tissue (T) and adjacent normal breast tissue (ANT); GAPDH was used as a loading control. (D) Real-time reverse transcription analysis of ETV4 mRNA expression in TNBC tissue and adjacent normal breast tissue; expression data were normalized to GAPDH. Error bars represent the mean ± standard deviation from three independent experiments; *P<0.05. (E) Immunohistochemistry staining of corresponding TNBC tissue (T) and adjacent normal breast tissue (ANT).Abbreviation: ER, estrogen receptor.

Mentions: To verify the array data, we performed Western blotting and real-time PCR analyses of ETV4 expression in TNBC cell lines and tissue specimens. We found that the expression of both ETV4 mRNA and protein was notably upregulated in hormone receptor-negative or TNBC cell lines tested (MDA-MB453, MDA-MB-231, and MDA-MB-435) compared with hormone receptor-positive cells (T47D, MDA-MB-415, MCF-7, ZR-75-30, BT474; Figure 2A and B). Furthermore, the data on eight pairs of TNBC tissue and adjacent normal breast tissue samples showed that expression of ETV4 mRNA and protein was also significantly higher in TNBC tissue than in the adjacent normal breast tissue (Figure 2C–E), indicating that ETV4 is overexpressed in TNBC cells and tissue. Also, results of IHC on eight pairs of tissue samples as described showed that ETV4 protein was positively expressed in TNBC tissue but not in the adjacent normal breast tissue.


Overexpression of ETV4 protein in triple-negative breast cancer is associated with a higher risk of distant metastasis.

Yuan ZY, Dai T, Wang SS, Peng RJ, Li XH, Qin T, Song LB, Wang X - Onco Targets Ther (2014)

Verification of ETS translocation variant 4 (ETV4) overexpression in triple-negative breast cancer (TNBC) cell lines and tissue.Notes: (A) Western blot analysis of ETV4 protein expression in TNBC cell lines and hormone receptor-positive cell lines. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B) Real-time reverse transcription analysis of ETV4 messenger ribonucleic acid (mRNA) expression in TNBC cell lines and hormone receptor-positive cell lines. Transcript levels were normalized to GAPDH. (C) Western blot analysis of ETV4 protein expression in TNBC tissue (T) and adjacent normal breast tissue (ANT); GAPDH was used as a loading control. (D) Real-time reverse transcription analysis of ETV4 mRNA expression in TNBC tissue and adjacent normal breast tissue; expression data were normalized to GAPDH. Error bars represent the mean ± standard deviation from three independent experiments; *P<0.05. (E) Immunohistochemistry staining of corresponding TNBC tissue (T) and adjacent normal breast tissue (ANT).Abbreviation: ER, estrogen receptor.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4196788&req=5

f2-ott-7-1733: Verification of ETS translocation variant 4 (ETV4) overexpression in triple-negative breast cancer (TNBC) cell lines and tissue.Notes: (A) Western blot analysis of ETV4 protein expression in TNBC cell lines and hormone receptor-positive cell lines. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B) Real-time reverse transcription analysis of ETV4 messenger ribonucleic acid (mRNA) expression in TNBC cell lines and hormone receptor-positive cell lines. Transcript levels were normalized to GAPDH. (C) Western blot analysis of ETV4 protein expression in TNBC tissue (T) and adjacent normal breast tissue (ANT); GAPDH was used as a loading control. (D) Real-time reverse transcription analysis of ETV4 mRNA expression in TNBC tissue and adjacent normal breast tissue; expression data were normalized to GAPDH. Error bars represent the mean ± standard deviation from three independent experiments; *P<0.05. (E) Immunohistochemistry staining of corresponding TNBC tissue (T) and adjacent normal breast tissue (ANT).Abbreviation: ER, estrogen receptor.
Mentions: To verify the array data, we performed Western blotting and real-time PCR analyses of ETV4 expression in TNBC cell lines and tissue specimens. We found that the expression of both ETV4 mRNA and protein was notably upregulated in hormone receptor-negative or TNBC cell lines tested (MDA-MB453, MDA-MB-231, and MDA-MB-435) compared with hormone receptor-positive cells (T47D, MDA-MB-415, MCF-7, ZR-75-30, BT474; Figure 2A and B). Furthermore, the data on eight pairs of TNBC tissue and adjacent normal breast tissue samples showed that expression of ETV4 mRNA and protein was also significantly higher in TNBC tissue than in the adjacent normal breast tissue (Figure 2C–E), indicating that ETV4 is overexpressed in TNBC cells and tissue. Also, results of IHC on eight pairs of tissue samples as described showed that ETV4 protein was positively expressed in TNBC tissue but not in the adjacent normal breast tissue.

Bottom Line: ETV4 messenger ribonucleic acid was more than five-fold upregulated in TNBC tissue compared with the control tissue.ETV4 overexpression was found in 57.0% of 135 TNBC cases.Overexpression of ETV4 protein was associated with an advanced stage and a higher proportion of positive lymph node and lymphovascular invasion.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncology in South China, Guangzhou, People's Republic of China ; Collaborative Innovation Center for Cancer Medicine, Guangzhou, People's Republic of China ; Department of Medical Oncology, Sun Yat-Sen University Cancer Center, Guangzhou, People's Republic of China.

ABSTRACT

Background: Patients with triple-negative breast cancer (TNBC) present a higher probability of distant metastasis and lack of effective targeted therapy. ETS translocation variant 4 (ETV4) is an ETS (E-26) transcription factor and has been associated with tumor metastasis. However, the clinical and functional significance of ETV4 in TNBC still remains unclear.

Methods: A human tumor metastasis polymerase chain reaction array was used to profile differential expression of tumor metastasis-related genes in TNBC tissue. Real-time reverse transcription and Western blot analyses were performed to verify ETV4 expression in TNBC cells and tissue. Immunohistochemistry was used to detect expression of ETV4 protein in 135 TNBC tissue samples for association between ETV4 protein expression and clinical outcomes.

Results: A total total of eight upregulated (CCL7, KISS1, MET, MMP7, NR4A3, ETV4, TIMP3, and TSHR) and three downregulated (ITGA7, SSTR, and MMP2) genes were identified between TNBC tissue and the luminal subtype of breast cancer tissue. ETV4 messenger ribonucleic acid was more than five-fold upregulated in TNBC tissue compared with the control tissue. ETV4 overexpression was found in 57.0% of 135 TNBC cases. Overexpression of ETV4 protein was associated with an advanced stage and a higher proportion of positive lymph node and lymphovascular invasion. Patients with an ETV4-overexpressed tumor had a significantly higher risk of developing distant metastasis (P<0.0001) and shorter overall survival and disease-free survival. Overexpression of ETV4 protein was an independent predictor of short disease-free survival of TNBC patients (P=0.021).

Conclusion: Overexpression of ETV4 protein increases risk of developing distant metastasis and results in a poor prognosis for TNBC patients. Thus, ETV4 might be a novel target for developing an alternative therapeutic strategy for prevention of TNBC distant metastasis.

No MeSH data available.


Related in: MedlinePlus