Assessment of the Cancer Risk of the Fat-Grafted Breast in a Murine Model.

Background: The results of experimental studies indicate that grafting of autologous adipose tissue may induce tumorigenesis at the recipient site, but clinical results do not support a carcinogenic effect of fat grafting to the breast. Objectives: The authors assessed cancer risk following transplantation of autologous fat into murine mammary tissue. Methods: In this animal study, mammary tissues from 54 breasts of 9 female rats were either grafted with autologous subcutaneous fat, grafted with autologous omental fat, or unmanipulated. Tissues were harvested and processed for histologic and immunohistochemical analyses, and the mRNA expression levels of specific genes were determined. Results: No atypia or changes in lobular structures were observed in lipofilled breasts compared with controls. The numbers of ductal cell layers and terminal ductal units were similar for lipofilled and control breasts. Macrophage concentrations also were similar for the 3 groups. The localization and magnitude of plasminogen activator inhibitor 1 were similar for lipofilled and unmanipulated breast tissue. The percentages of cells expressing Ki67 or estrogen receptor (ER) and the ER/Ki67 balance were similar for the 3 groups. Gene expression was not altered in lipofilled breasts, compared with controls. Conclusions: No theoretical risk of cancer was detected in the microenvironment of the lipofilled rat breast.

Prognostic significance of PD-L1 and PD-L2 in breast cancer.

PD-L1 and PD-L2 constitute an important antitumor immune response. In breast cancer, their prognostic value is still to be defined. In this study, we investigate the correlation between PD-L1 and PD-L2 protein expressions with clinical and pathologic features and disease-free survival and overall survival. To assess PD-L1 and PD-L2 expressions, we conducted immunohistochemistry studies using a breast cancer tissue microarray encompassing a total of 192 breast cancer cases, stages I, II, and III, with detailed clinical and outcome data. PD-L1 expression was present in 56.6% (107/189), and PD-L2 expression was identified in 50.8% (97/191) of breast cancer cases. Younger age at diagnosis, lymph node positivity, negative estrogen receptor, and recurrence at distant sites were all associated with both PD-L1 and PD-L2 expressions. The presence of larger tumors was associated only with PD-L1 expression. In our study, PD-L1 expression was significantly associated with better overall survival (P = .04) in breast cancer patients. Despite its association with poor clinical and pathologic features, PD-L1 expression emerges as a positive prognostic biomarker in breast cancer. This survival result might be due to the presence of a strong antitumor immune response leading to PD-L1 expression.

Unmanipulated native fat exposed to high-energy diet, but not autologous grafted fat by itself, may lead to overexpression of Ki67 and PAI-1.

BACKGROUND: Although its unclear oncological risk, which led to more than 20 years of prohibition of its use, fat grafting to the breast is widely used nowadays even for aesthetic purposes. Thus, we proposed an experimental model in rats to analyze the inflammatory activity, cellular proliferation and levels of Plasminogen Activator Inhibitor (PAI-1) in grafted fat, and in native fat exposed to high-energy diet in order to study the oncological potential of fat tissue. METHODS: Samples of grafted fat of rats on regular-energy diet were compared with paired samples of native fat from the same rat on regular-energy diet and on high-energy diet in a different time. Analysis involved microscopic comparisons using hematoxylin-eosin staining, immunohistochemistry with anti-CD68-labelled macrophages, and gene expression of Ki-67 and PAI-1. RESULTS: Hematoxylin-eosin staining analyses did not find any atypical cellular infiltration or unusual tissue types in the samples of grafted fat. The inflammatory status, assessed through immunohistochemical identification of CD68-labelled macrophages, was similar among samples of native fat and grafted fat of rat on regular-energy diet and of native fat of rats on high-energy diet. Real-time PCR revealed that high-energy diet, but not fat grafting, leads to proliferative status on adipose tissue (overexpression of ki-67, p = 0.046) and raised its PAI-1 levels, p < 0.001. CONCLUSION: While the native adipose tissue overexpressed PAI-1 and KI67 when exposed to high-energy diet, the grafted fat by itself was unable to induce cellular proliferation, chronic inflammatory activity and/or elevation of PAI-1 levels.

Is the immunohistochemical expression of proliferation (Ki-67) and apoptosis (Bcl-2) markers and cyclooxigenase-2 (COX-2) related to carcinogenesis in postmenopausal endometrial polyps?

OBJECTIVE: To evaluate the pattern of Ki-67, Bcl-2 and COX-2 expression in the glandular epithelium and stroma of malignant and benign endometrial polyps in postmenopause. STUDY DESIGN: A total of 390 postmenopausal women underwent surgical hysteroscopy; women with endometrial polyps were included. Polypoid lesions were histologically classified as benign, premalignant or malignant lesions. Ki-67, Bcl-2 and COX-2 expression were evaluated by immunohistochemistry according to percentage of stained cells, staining intensity, and final score. RESULTS: The prevalence of malignancy in endometrial polyps was 7.1% and was associated with postmenopausal bleeding. The final score showed that only mean COX-2 expression was higher in malignant polyps both in the glandular epithelium (6.1 +/- 2.5) (p < 0.001) and stroma (2.4 +/- 3.0) (p < 0.01). There was a higher Bcl-2 expression, especially in the glandular epithelium, with no differences between benign polyps and premalignant/malignant polyps. Ki-67 expression was low in both benign polyps and premalignant/malignant polyps. CONCLUSION: Polyps in postmenopause have a high COX-2 expression that is higher in malignant polyps than in benign polyps. There was no difference in Ki-67 and Bcl-2 expression between malignant polyps and premalignant/malignant polyps.

Prognostic significance of GRP78 expression patterns in breast cancer patients receiving adjuvant chemotherapy.

This study examined the associations between GRP78 expression and breast cancer recurrence and survival in patients treated with anthracyclines in the adjuvant setting. GRP78 expression was assessed in 106 stage II/III breast cancer patients. Tissue microarray was used to perform immunohistochemistry and to determine the GRP78 expression in endoplasmic reticulum and cell membrane of breast tumors. Four distinct scenarios (low and high thresholds) were developed. For high thresholds, 16% and 40% of our cases were GRP78-positive for endoplasmic reticulum and cell membrane, respectively. For low thresholds, 74% and 87% of our cases were GRP78-positive for endoplasmic reticulum and cell membrane, respectively. In the endoplasmic reticulum high-threshold scenario, GRP78 positive was found to be significantly frequent in T3 tumors (p=0.02), and inversely related to ERBB2 overexpression (p=0.03). There was a lower proportion of GRP78-positive cases among women between 50 and 65 years of age (p=0.02). In the endoplasmic reticulum low-threshold scenario, the proportion of GRP78-positive cases was significantly higher in women younger than 50 years and in those who were premenopausal (p=0.04). No statistically significant difference was found in survival probabilities among the scenarios examined. In our cohort, GRP78 overexpression was not a predictor of overall or disease-free survival of patients receiving anthracycline-based adjuvant chemotherapy.

The GSTT1 polymorphism of the glutathione S-transferase system in the intratumoral microvessel density of breast cancer patients.

It is well established that hypoxic microenvironment contributes to breast cancer progression by activation of transcriptional genes that promote angiogenesis. By promoting the antioxidant activity of glutathione, glutathione S-transferases (GSTs) are likely to facilitate the hypoxia-inducible factor-1α (HIF-1α) activity, therefore stimulating the angiogenesis. We investigated herein the influence of the GSTM1 and GSTT1 polymorphisms in the intratumoral angiogenesis of 87 patients with sporadic breast cancer. The intratumoral microvessel density (IMVD) of formalin-fixed paraffin-embedded tissues samples from all patients was determined by immunohistochemistry. The high IMVD was defined as a median microvessel counting higher than 18.7 after the analysis of histogram with all the results. The high IMVD was more common in patients with the GSTT1 wild genotype than in those with the GSTT1 null genotype (P = 0.04). Our results suggest, for the first time, that the GSTT1 polymorphism constitutes an inherited determinant of intratumoral angiogenesis in sporadic breast cancer.

The expression of the hormone receptors in the endometrium and endometrial polyps in postmenopausal women and its relationship to body mass index.

OBJECTIVE: To evaluate and compare the relationship of body mass index (BMI) and the immunoexpression of estrogen (ER) and progesterone receptors (PR) in endometrial polyps (EP) and endometrium, in gland and stromal cells of postmenopausal women. METHODS: Thirty-five postmenopausal women with benign endometrial polyps, who had not been taking medication with hormonal effects for at least 6 months, were submitted to operative hysteroscopy. The presence of ER and PR were evaluated by immunohistochemical method using a semiquantitative analysis. RESULTS: BMI was significantly higher among patients with lower expression of ER in the glands of endometrium (p=0.02). EP and adjacent endometrium showed significantly higher proportion of positive cells in the glands than in the stroma, for both ER (p=0.0015 and 0.0018, respectively) and PR (p=0.0176 and p<0.0001, respectively). Glands and stroma cells showed significantly higher proportion of positive cells in polyps than in the endometrium, for ER (p<0.0001 and p=0.0034, respectively). CONCLUSIONS: The higher proportion of positive gland cells for ER in EP as compared to endometrium supports an implication of these receptors in the pathogenesis of polyps. Association of higher BMI with lower expression of ER in endometrial glands, but not in EP, may indicate that factors influencing ER expression do not affect EP, supporting an autonomous function of polyps.

CD34 as a marker for evaluating angiogenesis in cervical cancer.

CD34 is an antigen present in hematopoietic progenitor cells and endothelial cells. Anti-CD34 antibody is a highly sensitive marker for endothelial cell differentiation and has also been studied as a marker for vascular tumors. However, there are few studies relating it to cervical carcinoma. The aim of this study was to evaluate the association between angiogenesis and the pathoanatomical features of cervical carcinoma using anti-CD34 monoclonal antibody. Sixty-two patients with invasive carcinoma of the uterine cervix in stages Ib-IIa were included. A primary monoclonal antibody specific for CD34 (Anti-Human Hemapoietic Progenitor Cell, CD34 Class II, Clone QBEnd 10, Code M7165, DAKO Corporation) was used in a dilution of 1:25. Microvessel density varied from 4.8 to 20 and was higher in undifferentiated carcinomas (p = 0.03; Mann-Whitney test). Higher microvessel density was associated with squamous cell carcinoma, odds ratio (OR) 8.8 (95% CI: 1.0-76.1), while the presence of lymphatic invasion yielded an OR of 2.6 (95% CI: 0.9-7.4). This study suggested that anti-CD34 antibody reactivity in cervical carcinoma is associated with pathoanatomical features indicative of poorer prognosis.

The genetics of hypertension modifies the renal cell replication response induced by experimental diabetes.

To investigate whether the genetics of hypertension modifies renal cell responses in experimental diabetes, we studied the renal cell replication and its regulation by two cyclin-dependent kinase (Cdk) inhibitors, p27(Kip1) and p21(Cip1), in prehypertensive spontaneously hypertensive rats (SHR) and their genetically normotensive counterparts, Wistar Kyoto (WKY) rats, with and without streptozotocin-induced diabetes. In diabetic SHR, the number of proliferating glomerular (0.6 +/- 0.3 positive cells/50 glomeruli) and tubulointerstitial (2.8 +/- 0.6 positive tubulointerstitial cells/50 grid fields) cells assessed by the bromodeoxyuridine technique was significantly (P = 0.0002) lower than in control SHR (13.2 +/- 1.7 and 48.6 +/- 9.7, respectively) and control (14.0 +/- 1.8 and 63.9 +/- 10.6) and diabetic (14.3 +/- 3.5 and 66.4 +/- 11.5) WKY rats. Proliferating cell nuclear antigen, another marker of cell proliferation, was significantly reduced in replicating glomerular (P = 0.0002) and tubulointerstitial (P < 0.0001) cells in diabetic SHR. In freshly isolated glomeruli, the level of p27(Kip1) detected by Western blotting was significantly higher in diabetic SHR than in nondiabetic SHR (1.52 +/- 0.14 vs. 1.00 +/- 0.10% of control, P = 0.014). The expression of p21(Cip1) in isolated glomeruli did not differ among the groups of rats. In conclusion, the response of renal cell replication to diabetes differs markedly between prehypertensive SHR and their WKY control rats. The decreased glomerular cell proliferation in prehypertensive diabetic SHR is at least partly mediated by a higher expression of the Cdk inhibitor p27(Kip1).