Effects of integrin av ß3 inhibitor CycIo-RGDfK on epithelial- mesenchymal transition of breast epithelial cells induced by fibronectin / 中国药理学通报

Aim To investigate the effect of integrin av ß3 inhibitor (Cyclo-RGDfK) on fibronectin (F N) - induced epithelial-mesenchymal transition in human mammary epithelial cells MCF-10A. Methods M C F - 10A cells were cultured in vitro, and the model of epithelial-mesenchymal transition (E M T) induced by FN was established. The expressions of otv integrin, ß3 integrin, epithelial marker E-cadherin and mesenchymal markers N-cadherin and Vimentin were measured by Western blot. The migration ability and invasion ability of cells were detected by Scratch repair experiment and Transwell chamber experiment. Results Western blot results showed that the expressions of ay integrin, ß3 integrin, N-cadherin, and Vimentin significantly were up-regulated and the expression of E-cadherin protein down-regulated after FN treatment for 48 h. After administration of Cyclo-RGDfK, the increased expression of N-cadherin and Vimentin and decreased expression of E-cadherin induced by FN were significantly reversed. Similarly, the results of Scratch repair experiments and Transwell experiments showed that FN significantly enhanced the ability of MCF-10A cells to migrate and invade, and Cyclo-RGDfK significantly inhibited the induction of FN. Conclusions Cyclo-RGDfK can inhibit the process of EMT induced by FN and reduce the invasion and migration of MCF-10A, which may be a potential drug for the treatment of breast cancer metastasis.

Can widely used cell type markers predict the suitability of immortalized or primary mammary epithelial cell models?

BACKGROUND: Mammary cell cultures are convenient tools for in vitro studies of mammary gland biology. However, the heterogeneity of mammary cell types, e.g., glandular milk secretory epithelial or myoepithelial cells, often complicates the interpretation of cell-based data. The present study was undertaken to determine the relevance of bovine primary mammary epithelial cells isolated from American Holstein (bMEC US) or Swiss Holstein-Friesian (bMEC CH) cows, and of primary bovine mammary alveolar epithelial cells stably transfected with simian virus-40 (SV-40) large T-antigen (MAC-T) for in vitro analyses. This was evaluated by testing their expression pattern of cytokeratin (CK) 7, 18, 19, vimentin, and α-smooth muscle actin (α-SMA. RESULTS: The expression of the listed markers was assessed using real-time quantitative PCR, flow cytometry and immunofluorescence microscopy. Characteristic markers of the mesenchymal (vimentin), myoepithelial (α-SMA) and glandular secretory cells (CKs) showed differential expression among the studied cell cultures, partly depending on the analytical method used. The relative mRNA expression of vimentin, CK7 and CK19, respectively, was lower (P < 0.05) in immortalized than in primary mammary cell cultures. The stain index (based on flow cytometry) of CK7 and CK19 protein was lower (P < 0.05) in MAC-T than in bMECs, while the expression of α-SMA and CK18 showed an inverse pattern. Immunofluorescence microscopy analysis mostly confirmed the mRNA data, while partly disagreed with flow cytometry data (e.g., vimentin level in MAC-T). The differential expression of CK7 and CK19 allowed discriminating between immortal and primary mammary cultures. CONCLUSIONS: The expression of the selected widely used cell type markers in primary and immortalized MEC cells did not allow a clear preference between these two cell models for in vitro analyses studying aspects of milk composition. All tested cell models exhibited to a variable degree epithelial and mesenchymal features. Thus, based on their characterization with widely used cell markers, none of these cultures represent an unequivocal alveolar mammary epithelial cell model. For choosing the appropriate in vitro model additional properties such as the expression profile of specific proteins of interest (e.g., transporter proteins) should equally be taken into account.

Expression of maspin in benign and malignant salivary gland tumors: An immunohistochemical study.

Context: Maspin is a novel serine protease inhibitor (serpin) with multifaceted tumor‑suppressive activities. It was originally identified in normal human breast myoepithelial cells and shows variable expression in different types of cancer cells. Maspin displays anti‑metastatic properties in mammary and prostate cancer. Its expression is maintained during ovarian, lung and pancreatic carcinogenesis, indicating that Maspin regulated metastatic potential is tissue specific. Thus, it is possible that Maspin participates in salivary gland tumor biology as well. In this study, expression pattern of maspin in benign and malignant salivary gland tumors is analyzed, to understand the biological behavior of salivary gland tumors with respect to maspin expression. Aims and Objectives: The aim of this study was to demonstrate, record, and correlate the expression pattern of maspin in benign and malignant salivary gland tumors. Settings and Design: A retrospective study of maspin expression in 30 diagnosed cases of benign and malignant salivary gland tumors retrieved from archives of our department. Materials and Methods: Anti‑maspin antibody and horseradish peroxidase detection system. Statistical Analysis: Descriptive statistical analysis and Chi‑square/Fisher Exact test. Results: Intense expression with P < 0.001 is associated with benign tumors, nuclear staining with P < 0.001 is significantly associated with benign tumors and cytoplasmic staining with P = 0.020 is associated with malignant tumors. Conclusion: Intensity of expression is more in benign tumors when compared with malignant tumors. The benign tumors showed both nuclear and cytoplasmic expression. Some malignant tumors did express maspin, but mainly in the cytoplasm.

HIV transmission through breastmilk: the science behind the understanding of current trends and future research

Breastmilk protects the infant from many diseases and many short- term and long- term benefits accrue. At the same time it is also known that breastfeeding acts as a vehicle for some infective agents. It is now accepted that breastmilk transmission of Human Immunodeficiency Virus- 1 (HIV-1) is an important mode of paediatric infection . Despite this fact, many researchers have observed that corresponding to the volume of milk consumed by the infant, maternal transmission via breastmilk is still comparatively low. Some have noted the long latency period of breastmilk HIV transmission with evidence of numerous anti-HIV factors in breastmilk. Although there are accepted standard guidelines on infant feeding in mothers who are HIV positive in many countries, it maybe equally important to realize gaps in our knowledge of mother- to -child HIV transmission. From an evolutionary perspective, the role of the mammary epithelial cell (MEC) and of breastmilk , in contributing to and possibly in influencing HIV-1 transmission is intriguing. The presence of HIV-1 or of other viruses in maternal milk seem to be a requisite to spur immunological defenses to optimize necessary protection to the infant. This article reviews some aspects of the science of HIV transmission through breastmilk and reflects the concept -based understanding of current policies on HIV and breastfeeding. At the same time, it highlights uncertainties in this field and the urgency for future research in this direction. Accepting current notions of breastmilk HIV transmission, greater deliberation by research may throw more light on why breastfeeding with its abundant advantages is fraught with the hazards of transmission of a deadly disease.

miR-138 Function and Its Targets on Mouse Mammary Epithelial Cells / 生物化学与生物物理进展

miRNA was 20～25 nt endogenous non-coding RNA. miRNAs are encoded small RNAs that hybridize with messenger RNAs, resulting in degradation or translational inhibition of targeted transcripts. In order to investigate the function of miR-138 on mouse mammary epithelial cells, technique for gene silencing-miRNA inhibitor (anti-miRNA) was applied to make miR-138 silence, qRT-PCR was showed valid for inhibitor miR-138. And Western blot, CASY(r)-technology was put in use to study some change of mouse mammary epithelial cells after miR138 inhibitor. It was shown that miR-138 suppresses the exepress of PRL-R(P

Primary culture of nontumorigenic human mammary epithelial cells / 基础医学与临床

Objective To investigate the feasibility of primary culture of nontumorigenic human mammary epithelial cells,and the effect of proliferation due to hormone.Methods Normal human mammary tissue was taken from mammary segment ectomy,and the tissue was digested by collagenase type Ⅰ.Epithelial depuration was carried out by integration of burning and enzyme digestion.The cells we acquired were identified by observation of cell morphology by light and Transmission Electron Microscopy(TEM).Results It is feasible to culture nontumorigenic human mammary epithelial cells.?-estradiol and progesterone at 1?10-5 g/L can promote the proliferation of the normal human mammary epithelial cells.