J Breast Cancer.  2017 Mar;20(1):35-44. 10.4048/jbc.2017.20.1.35.

MicroRNA-222 Expression as a Predictive Marker for Tumor Progression in Hormone Receptor-Positive Breast Cancer

• Han SH
• Kim HJ
• Gwak JM
• Kim M
• Chung YR
• Park SY

• KMID: 2379397
• DOI: http://doi.org/10.4048/jbc.2017.20.1.35

Abstract

PURPOSE
The microRNA-221/222 (miR-221/222) gene cluster has been reported to be associated with the promotion of epithelial-mesenchymal transition (EMT), downregulation of estrogen receptor-α, and tamoxifen resistance in breast cancer. We studied the expression of miR-222 in human breast cancer samples to analyze its relationship with clinicopathologic features of the tumor, including estrogen receptor status, expression of EMT markers, and clinical outcomes.
METHODS
Quantitative real-time polymerase chain reaction was performed to detect the expression of miR-222 in 197 invasive breast cancers. Expression of EMT markers (vimentin, smooth muscle actin, osteonectin, N-cadherin, and E-cadherin) was evaluated using immunohistochemistry.
RESULTS
High miR-222 levels were associated with high T stage, high histologic grade, high Ki-67 proliferation index, and HER2 gene amplification. Its expression was significantly higher in the luminal B and human epidermal growth factor receptor 2-positive (HER2+) subtypes than in the luminal A and triple-negative subtypes. In the hormone receptor-positive subgroup, there was a significant negative correlation between miR-222 and estrogen receptor expression, and miR-222 expression was associated with EMT marker expression. In the group as a whole, high miR-222 expression was not associated with clinical outcome. However, subgroup analyses by hormone receptor status revealed that high miR-222 expression was a poor prognostic factor in the hormone receptor-positive subgroup, but not in the hormone receptor-negative subgroup.
CONCLUSION
This study showed that miR-222 is associated with down-regulation of the estrogen receptor, EMT, and tumor progression in hormone receptor-positive breast cancer, indicating that miR-222 might be associated with endocrine therapy resistance and poor clinical outcome in hormone receptor-positive breast cancer.

Keyword

Breast neoplasms; Epithelial-mesenchymal transition; miR-222; Prognosis

MeSH Terms

Actins
Breast Neoplasms*
Breast*
Cadherins
Down-Regulation
Epithelial-Mesenchymal Transition
Estrogens
Genes, erbB-2
Humans
Immunohistochemistry
Multigene Family
Muscle, Smooth
Osteonectin
Phenobarbital
Prognosis
Real-Time Polymerase Chain Reaction
Receptor, Epidermal Growth Factor
Tamoxifen
Actins
Cadherins
Estrogens
Osteonectin
Phenobarbital
Receptor, Epidermal Growth Factor
Tamoxifen

Figure

• 

  Figure 1 Representative examples of immunohistochemical staining (IHC) for epithelial-mesenchymal transition marker in breast cancers. (A) Diffuse expression of vimentin with score of 5 (IHC for vimentin, ×200). (B) Scattered expression of osteonectin with score of 4 (IHC for osteonectin, ×200). (C) Focal expression of N-cadherin with score of 3 IHC for N-cadherin (IHC for N-cadherin, ×200).

• 

  Figure 2 Difference in microRNA-222 (miR-222) expression levels according to breast cancer subtypes. The box shows the first to third quartiles of miRNA expression levels, the horizontal line inside the box represents the median, the whiskers extend to minimum and maximum values within 1.5 times the interquartile range (IQR) from the first and third quartiles. Outliers are represented by small circles and, extreme values (more than 3 times IQR), by asterisks. miR-222 expression is higher in luminal B (LB) and human epidermal growth factor receptor 2-positive (HER2+) subtypes than in luminal A (LA) and triple-negative (TN) subtypes.

• 

  Figure 3 Correlation between microRNA-222 (miR-222) and estrogen receptor (ER) expression level. In hormone receptor-positive subgroup, miR-222 expression level shows a mild, but significant negative correlation with estrogen receptor expression level.

• 

  Figure 4 Epithelial-mesenchymal transition (EMT) marker expression in relation to microRNA-222 (miR-222) expression level. In whole study group, vimentin expression is higher in tumors with low level of miR-222 expression (A). While there are no correlations between miR-222 expression levels and EMT marker expression in triple-negative breast cancer (TNBC) subgroup (B), N-cadherin expression and E-cadherin loss are more frequent in tumors with high level of miR-222 expression in non-TNBC (C).SMA=smooth muscle actin. *Statistically significant difference.

• 

  Figure 5 Disease-free survival according to microRNA-222 (miR-222) expression levels. (A) whole study population, (B) hormone receptor-positive subgroup, (C) hormone receptor-negative subgroup. Patients with high levels of miR-222 expression show significantly shorter disease-free survival time compared to those with low levels of miR-222 in hormone receptor-positive subgroup (B), but not in whole study population (A) and hormone receptor-negative subgroup (C).

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