Human Malignant Breast Cell Research Articles

The Secretome of Human Neonatal Mesenchymal Stem Cells Modulates Doxorubicin-Induced Cytotoxicity: Impact in Non-Tumor Cells.

Doxorubicin (Dox) is one of the most widely used treatments for breast cancer, although limited by the well-documented cardiotoxicity and other off-target effects. Mesenchymal stem cell (MSC) secretome has shown immunomodulatory and regenerative properties, further potentiated under 3D conditions. This work aimed to uncover the effect of the MSC-derived secretome from 3D (CM3D) or 2D (CM2D) cultures, in human malignant breast cells (MDA-MB-231), non-tumor breast epithelial cells (MCF10A) and differentiated AC16 cardiomyocytes, co-treated with Dox. A comprehensive proteomic analysis of CM3D/CM2D was also performed to unravel the underlying mechanism. CM3D/CM2D co-incubation with Dox revealed no significant differences in MDA-MB-231 viability when compared to Dox alone, whereas MCF10A and AC16 viability was consistently improved in Dox+CM3D-treated cells. Moreover, neither CM2D nor CM3D affected Dox anti-migratory and anti-invasive effects in MDA-MB-231. Notably, Ge-LC-MS/MS proteomic analysis revealed that CM3D displayed protective features that might be linked to the regulation of cell proliferation (CAPN1, CST1, LAMC2, RANBP3), migration (CCN3, MMP8, PDCD5), invasion (TIMP1/2), oxidative stress (COX6B1, AIFM1, CD9, GSR) and inflammation (CCN3, ANXA5, CDH13, GDF15). Overall, CM3D decreased Dox-induced cytotoxicity in non-tumor cells, without compromising Dox chemotherapeutic profile in malignant cells, suggesting its potential use as a chemotherapy adjuvant to reduce off-target side effects.

Regulation of Cell- Generated Forces during Phenotypic Reversion by a Novel Laminin Chain in 3D Culture of Human Breast Tumor Cells

Laminins are heterotrimeric extracellular glycoproteins found in, but not confined to, basement membranes (BMs). They are important components in formation of the molecular networks of BMs as well as in cell & tissue polarity, differentiation and morphogenesis. They are secreted by cells and serve to initiate the template required for multicellular assembly. Mass spectroscopy of the isolated membrane fractions revealed a differential expression of an unexpected laminin chain where human malignant breast cells have a higher expression than that of non malignant cells in 3D lrECM. We thus asked the question what is the role of this endogenously produced laminin for 3D acinar morphogenesis. Recently, we reported that non-malignant cells in a 3D laminin-rich gel generate a centripetal force resulting in coherent angular motion (CAMo) to establish acini. On the other hand, malignant cells are randomly motile but regain the ability to rotate during ‘phenotypic reversion'. We show that there is a differential deposition of this laminin in the BM in malignant tissues from human patient biopsies. Modulation of expression via shRNA reveals that malignant cells regain the ability to generate centripetal forces and thus re-enter the acinar morphogenetic program. We determine that the cells' ability to perform CAMo is in part due to both the expression and localization of this particular laminin chain. These data show the intimate dynamic reciprocity between the cells and the ECM to establish polarity and form tissue architecture

Effects of extracellular YB-1 protein on cultured cells of human breast cancer

Multifunctional protein YB-1, which takes a part in many DNA/RNA-associated events in cells (transcription, mRNA translation, splicing) is a promising prognostic and predictive marker for various human tumors. Recently it was discovered that protein YB-1 secreted by cells functions as an extracellular mitogen. These results were obtained on kidney mesangial cells and monocytes. Here we studied the effects of extracellular YB-1 on the cell number, migration and gene expression in the populations of three lines of human malignant breast cells (MCF-7, HBL-100, BT-474). We used recombinant full-length protein YB-1, which retained all the functional sites necessary for its various activities, including binding to the receptor Notch-3 (rYB-1). Extracellular rYB-1 increased the number of cells 2- to 3-fold in the tested lines. The wound was made in cell monolayers of HBL-100, MCF-7 and MCF-7/ras. Protein rYB-1 increased the rate of wound healing in MCF-7/ras. Sensitivity to chemotherapeutic agents did not change in the presence of rYB-1 in any of the cell lines studied. We have used expression microarrays to study the effects of rYB-1 on the profile of gene expression in MCF-7 cells and in MCF-7 cells transfected with YB-1-GFP construct. In MCF-7 cells extracellular rYB-1 mainly down-regulated different genes, while in the cells transfected with cDNA of YB-1-GFP the activity of different genes was up-regulated. Our results show that extracellular YB-1 protein and intracellular YB-1 produce diverse changes in the cells of mammary tumors. It is possible that extracellular YB-1 is involved in the regulation of cell proliferation and migration in the metastatic niche.

Synthesis of some new diaryl and triaryl hydrazone derivatives as possible estrogen receptor modulators.

1,2-Bis(4-substituted phenyl)-2-methyl ethanone (2,4-dinitrophenyl)hydrazones and 1-naphthyl-1-(4-substituted phenyl)-methanone (2,4-dinitrophenyl)hydrazones have been synthesized and evaluated for their anti-implantation, uterotrophic, antiuterotrophic, anticancer and antimicrobial activities. Diphenolic hydrazone (compound 6) showed maximum uterotrophic inhibition of 70%, whereas compound 20 exhibited cytotoxicity in the range of 50-70% against MCF-7 and ZR-75-1 human malignant breast cell lines.