Breast Cancer/Stromal Cells Coculture on Polyelectrolyte Films Emulates Tumor Stages and miRNA Profiles of Clinical Samples

Abstract

In this study, we demonstrate a method for controlling breast cancer cells adhesion on polyelectrolyte multilayer (PEM) films without the aid of adhesive proteins/ ligands to study the role of tumor and stromal cell interaction on cancer biology. Numerous studies have explored engineering coculture of tumor and stromal cells predominantly using transwell coculture of stromal cells cultured onto coverslips that were subsequently added to tumor cell cultures. However, these systems imposed an artificial boundary that precluded cell−cell interactions. To our knowledge, this is the f irst demonstration of patterned coculture of tumor cells and stromal cells that captures the temporal changes in the miRNA signature as the breast tumor develops through various stages. In our study we used synthetic polymers, namely poly(diallyldimethylammonium chloride) (PDAC) and sulfonated poly(styrene) (SPS), as the polycation and polyanion, respectively, to build PEMs. Breast cancer cells attached and spread preferentially on SPS surfaces while stromal cells attached to both SPS and PDAC surfaces. SPS patterns were formed on PEM surfaces, by either capillary force lithography (CFL) of SPS onto PDAC surfaces or vice versa, to obtain patterns of breast cancer cells and patterned cocultures of breast cancer and stromal cells. In this study, we utilized cancer cells derived from two different tumor stages and two different stromal cells to effectively model a heterogeneous tumor microenvironment and emulate various tumor stages. The coculture model mimics the proliferative index (Ki67 expression) and tumor aggressiveness (HER-2 expression) akin to those observed in clinical tumor samples. We also demonstrated that our patterned coculture model captures the temporal changes in the miRNA-21 and miRNA-34 signature as the breast tumor develops through various stages. The engineered coculture platform lays groundwork toward precision medicine wherein patient-derived tumor cells can be incorporated within our in vitro models to identify potential pathways and drug treatment regimens for individual patients.

Depletion of mitochondrial protease OMA1 alters proliferative properties and promotes metastatic growth of breast cancer cells

“…5A). Ki67, a nuclear protein, is a commonly used clinical marker to determine the proliferative state of tumor cells 3,4,40,41 . Remarkably, the Ki67 expression was higher in the OM.21 cells, indicating that these cells are in a hyperproliferative state relative to normal 21MT-1 cells (Fig.…”

MicroRNA-34 Family in Cancers: Role, Mechanism, and Therapeutic Potential

“…Furthermore, miR-34 mimics have been shown to sensitize CSCs to chemotherapy alone. A combination of miR-34a mimics and doxorubicin, a commonly used chemotherapy drug used in breast cancer, synergistically inhibited breast cancer CSC properties and significantly reduced tumor growth compared to either treatment alone by sensitizing the CSCs to doxorubicin-induced apoptosis, thereby overcoming chemoresistance [151,152].…”

“…Thymoquinone (TQ, a derivative of the Chinese herbal medicine dendrobium) possesses potent antitumor properties, including the inhibition of cell proliferation and induction of apoptosis. When combined with miR-34 mimics, overall better treatment outcomes were observed [152,185]. Another natural compound with anticancer properties is curcumin.…”

Sistema de cocultura com as linhagens celulares humanas HepG2 e HUVEC na investigação da genotoxicidade de toxinas isoladas de Bothrops jararacussu

“…Uma das maiores dificuldades é reproduzir o microambiente tumoral utilizando apenas uma linhagem celular, então, sistemas de coculturas utilizando pelo menos duas linhagens celulares estão sendo estabelecidos para elucidar as interações célula-célula e célula-microambiente tumoral (MIKI et al, 2012;DAVEREY et al, 2015). A densidade de células HepG2 e HUVEC cultivadas nos insertos foi baseada em estudos anteriores.…”

“…Este processo ocorre durante o desenvolvimento normal e envelhecimento como um mecanismo fisiológico essencial para manutenção da homeostase em populações de células nos tecidos e, além disso, pode ocorrer como um mecanismo de defesa em reações imunológicas ou quando as células sofrem danos causados por agentes nocivos ou por doenças(NORBURY e HICKSON, 2001). meio de fatores de crescimento tumorais, citocinas e quimiocinas(FANG e DECLERCK, 2013;BOELENS et al, 2014;DAVEREY et al, 2015). Para isso, técnicas in vitro envolvendo a cultura simultânea de mais de uma linhagem celular, permitindo o compartilhamento do meio de cultura e, assim, mantendo a comunicação parácrina entre essas diferentes células, têm sido empregadas.…”