Background: The search for new markers for breast cancer (BC) has been sought in order to better understand this type of cancer. The Heart and Neural Crest Derivatives Expressed Transcript 2 (HAND2) has been related to endometrial cancer but there are scant data related to BC. The aim of this study is to compare the immunohistochemical expression of HßAND2 in normal breast tissue vs. BC and to correlate with the estrogen receptor (ERα). Patients and Methods: In this case-control study, 19 formalin–fixed, paraffin-embedded tissues were obtained from pathological archives. Benign (n=9; control) and cancer (n=10) breast tissue were analyzed with immunohistochemistry for HAND2 (Ab60037), at dilution 1:50 at pH 9 and ERα (SP1). ImageJ software with "color deconvolution" was used for analysis of the expression of these proteins. The sample size was calculated (power=95%, α error= 1%) to identify an increase from mean 15 DAB units (control) to 40 DAB units in cancer. Results: HAND2 expression (mean ± SD) was 15.5 ± 6.1 (cancer) versus 44.8 ± 21.1 (control) (P=0.002, Student t-test). Its expression was mainly present in the cytosol of the cells. No correlation was observed between ERα and HAND2 (Pearson r = -0.28 (95%CI=-0.6 to 0.22; P=0.2). Conclusions: The protein expression of HAND2, using Ab60037 antibody, is reduced in breast cancer, compared with normal breast tissue. The expression of HAND2 is not correlated with ERα expression.

Objectives: To verify the immunohistochemical expression of Heart and Neural Crest Derivatives Expressed Transcript 2 (HAND2), a tumor suppressor protein, .in breast tissues with and without breast cancer. To correlate the expression of HAND2 with estrogen receptor (ER). Methods: In this case-control study, 19 formalin-fixed, paraffin-embedded tissues were obtained for pathological archives analysis. Tissue sections of breast tissue derived from benign conditions (n=10) and breast cancer (n=9) were analyzed for the intensity of the staining with 3,3'-diaminobenzidine using rabbit polyclonal antibody against HAND2 (Ab60037), at dilution 1:50 at pH 9; ER was analyzed using clone 1D5, monoclonal, Dako) diluído a 1/100. ImageJ software with "color deconvolution" was used for analysis of the expression of these proteins. Statistical analysis was performed using unpaired t-test and correlation of Pearson. The sample size was calculated in order to have a power of 95%, an alpha error of 1% to identify an increase in the primary outcome measure from 15 in the control group to 44 in the experimental group. Results: HAND2 expression (mean±SD) in the breast tissue was 15.5±1.9 (cancer) versus 44.9±7 (breast cancer) (p=0.0006). Its expression was present in the nuclear and cytoplasmic compartments of the cells. No correlation was observed between ER and HAND2 (Pearson r =-0.28 (95%CI-0.6-0.22; p=0.2). Conclusions: The immunoexpression of HAND2 is reduced in breast cancer, compared with normal breast tissue. The expression of HAND2 is not correlated to ER expression These findings may guarantee further research as a potential independent prognostic biomarker.

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