T-type Ca2+ channels and their relationship with pre-neoplastic and neoplastic lesions in the human breast.

The expression of T-type voltage-dependent Ca2+ channels (Cav3) has been previously observed in breast cancer, but their expression and subcellular localization were not evaluated in pre-neoplastic lesions. Therefore, this work aimed to evaluate protein expression and subcellular localization of T-type channel isoforms in human breast tissue samples. Protein expressions of CaV3.1, CaV3.2, and CaV3.3 were evaluated by immunohistochemistry in breast without alteration, in proliferative non-neoplastic lesions, and in neoplastic ductal epithelial lesions of the human breast. CaV3.1, CaV3.2, and CaV3.3 nuclear expressions were decreased in advanced stages of neoplastic transformation, whereas CaV3.1 and CaV3.2 cytoplasmic expression increased. Also, the decrease in nuclear expression was correlated with an increase in cytoplasmic expression for CaV3.1 isoform. The change in CaV3 protein expression and subcellular localization are consistent with the neoplastic transformation stages of mammary epithelial cells, evident in early neoplastic lesions, such as ductal carcinomas in situ. These results suggest a possible involvement of CaV3 in the carcinogenic processes and could be considered as a potential pharmacological target in new therapies for breast cancer treatment.

T-type Ca2+ channels and their relationship with pre-neoplastic and neoplastic lesions in the human breast

The expression of T-type voltage-dependent Ca2+ channels (Cav3) has been previously observed in breast cancer, but their expression and subcellular localization were not evaluated in pre-neoplastic lesions. Therefore, this work aimed to evaluate protein expression and subcellular localization of T-type channel isoforms in human breast tissue samples. Protein expressions of CaV3.1, CaV3.2, and CaV3.3 were evaluated by immunohistochemistry in breast without alteration, in proliferative non-neoplastic lesions, and in neoplastic ductal epithelial lesions of the human breast. CaV3.1, CaV3.2, and CaV3.3 nuclear expressions were decreased in advanced stages of neoplastic transformation, whereas CaV3.1 and CaV3.2 cytoplasmic expression increased. Also, the decrease in nuclear expression was correlated with an increase in cytoplasmic expression for CaV3.1 isoform. The change in CaV3 protein expression and subcellular localization are consistent with the neoplastic transformation stages of mammary epithelial cells, evident in early neoplastic lesions, such as ductal carcinomas in situ. These results suggest a possible involvement of CaV3 in the carcinogenic processes and could be considered as a potential pharmacological target in new therapies for breast cancer treatment.

Comparative analysis of six different antibodies against Her2 including the novel rabbit monoclonal antibody (SP3) and chromogenic in situ hybridisation in breast carcinomas.

AIMS: To compare the sensitivity and specificity of new rabbit monoclonal antibody SP3 with those of mouse monoclonal and rabbit polyclonal antibodies using HER2 amplification defined by chromogenic in situ hybridisation (CISH) as the gold standard. METHODS: Serial sections from tissue microarrays (TMAs) containing 84 breast carcinomas were submitted to CISH (Zymed HER2 Spot-Light kit) and immunohistochemistry, using NeoMarkers SP3 (rabbit monoclonal), DAKO A0485 and DAKO HercepTest (polyclonal), Novocastra NCL-CB11, Cell Marque CM-CB11, and Genentech 4D5 (mouse monoclonal). RESULTS: The best antibody concordance was between SP3 and HercepTest (kappa = 0.74). SP3, A0485 and HercepTest detected all HER2 amplified tumours, but were less specific than mouse monoclonal antibodies. 3/38 (7.9%) and 8/38 (21.0%) non-amplified tumours were scored as 3+ using SP3 and A0485, respectively. 3/46 (6.5%) amplified tumours were negative for NCL-CB11. SP3, HercepTest and A0485 showed no gene amplification on 55%, 62.5% and 92.3% of the 2+ scored tumours, but most of the 2+ scored tumours using monoclonal antibodies were amplified by CISH (80-92.3%). CONCLUSIONS: SP3 is more sensitive than mouse monoclonal antibodies for Her2 assessment. However, HercepTest, CB11 and 4D5 show higher specificity than SP3 for the identification of HER2 gene amplification. Mouse monoclonal antibodies show less Her2 2+ tumours; most are amplified by CISH.

Morphology and healing resistance of abdominal wall after longitudinal and transversal laparotomy in rabbits.

AIM: The abdominal wall continues to be a topic of investigation for the evaluation of its healing in terms of morphology and resistance. In the present investigation, transverse and longitudinal laparotomies were studied comparatively. METHODS: Thirty rabbits were divided into two groups: Group 1 (n=10) longitudinal laparotomy, Subgroup 1A (n=5) suture of the anterior and posterior sheaths of the abdominal rectus muscle and of the peritoneum, Subgroup 1B (n=5) suture of the anterior sheath of the abdominal rectus muscle; Group 2 (n=20) transverse laparotomy, Subgroup 2A (n=5) suture of the anterior and posterior sheaths of the abdominal rectus muscle and of the peritoneum, Subgroup 2B (n=5) suture of only the anterior sheath of the abdominal rectus muscle, Subgroup 2C (n=5) suture of the abdominal rectus muscle and of its anterior sheath on a single plane, Subgroup 2D (n=5) repair of the posterior sheath of the abdominal rectus muscle together with the peritoneum, followed by suture of the abdominal rectus muscle complemented with suture of the anterior sheath of the same muscle. After 17 days, two peritoneal aponeurotic muscular segments of the scar were removed for the evaluation of resistance and of histological aspects. RESULTS: The resistance values detected for each group showed 1A>1B, 1A>2A and 1B>2B, and 2B>2C>2D>2A (P=0.014). Dehiscence, infections and adhesions were more frequent in Group 2. Histology revealed muscular degeneration and necrosis, with mature fibrous connective scar tissue replacing muscle tissue. CONCLUSION: Transverse muscle section causes greater muscle weakening and leaving the peritoneum open does not alter the resistance of the scar.