Dear Hanie,

Attached please find a paper that covers the answer to your question. I have copied some of the methods for quick view.

Research articleMesenchymal stem cells mediate the clinical phenotype of inflammatory breast cancer in a preclinical model

Lara Lacerda12, Bisrat G Debeb12, Daniel Smith12, Richard Larson12, Travis Solley1, Wei Xu12, Savitri Krishnamurthy23,Yun Gong23, Lawrence B Levy1, Thomas Buchholz12, Naoto T Ueno24, Ann Klopp1† and Wendy A Woodward12*†

Breast Cancer Research 2015, 17:42 doi:10.1186/s13058-015-0549-4

Abstract

Introduction

Inflammatory breast cancer (IBC) is an aggressive type of breast cancer, characterized by very rapid progression, enlargement of the breast, skin edema causing an orange peel appearance (peau d’orange), erythema, thickening, and dermal lymphatic invasion. It is characterized by E-cadherin overexpression in the primary and metastatic disease, but to date no robust molecular features that specifically identify IBC have been reported. Further, models that recapitulate all of these clinical findings are limited and as a result no studies have demonstrated modulation of these clinical features as opposed to simply tumor cell growth.

Methods

Hypothesizing the clinical presentation of IBC may be mediated in part by the microenvironment, we examined the effect of co-injection of IBC xenografts with mesenchymal stem/stromal cells (MSCs).

Results

MSCs co-injection significantly increased the clinical features of skin invasion and metastasis in the SUM149 xenograft model. Primary tumors co-injected with MSCs expressed higher phospho-epidermal growth factor receptor (p-EGFR) and promoted metastasis development after tumor resection, effects that were abrogated by treatment with the epidermal growth factor receptor (EGFR) inhibitor, erlotinib. E-cadherin expression was maintained in primary tumor xenografts with MSCs co-injection compared to control and erlotinib treatment dramatically decreased this expression in control and MSCs co-injected tumors. Tumor samples from patients demonstrate correlation between stromal and tumor p-EGFR staining only in IBC tumors.

Conclusions

Our findings demonstrate that the IBC clinical phenotype is promoted by signaling from the microenvironment perhaps in addition to tumor cell drivers.

Materials and methods

Cell culture

The IBC cell line SUM149 was obtained from Asterand (Detroit, MI, USA) and cultured in Ham’s F-12 media supplemented with 10% fetal bovine serum (FBS), 1 mg/mL hydrocortisone, 5 mg/mL insulin, and 1% antibiotic-antimycotic. Human-derived bone marrow MSCs were obtained from EMD Millipore (Billerica, MA, USA) (Part #SCC034, Lot N61710996) and cultured in alpha minimum essential medium (MEM) supplemented with 20% FBS and 1% penicillin/streptomycin/glutamine.

Hoping this will be helpful,

Rafik

Yes, indeed there are a number of breast cancer cell lines that can be classified as mesenchymal-like. See this paper:

Lehmann BD, Bauer JA, Chen X, Sanders ME, Chakravarthy AB, Shyr Y, Pietenpol JA. J Clin Invest. 2011 Jul;121(7):2750-67.

They classified 2 types of mesenchymal-like breast cancers a Mesenchymal group (M) and a mesenchymal stem-like group (MSL) in primary breast cancers and also assigned a variety of cell lines to these groups: CAL-120, CAL-51, MDA-MB-157, MDA-MB-231, MDA-MB-436, SUM159PT, HS578T and BT549.

MDA-MB-231 are probably the most commonly used.

Good luck,

Phil

Dear Rafik and Philip, Thank you for your responses!

Dr. Coates used "mesenchymal-like" phrase. What does that means? I always thought that the life cycle of cancer cells are as follow:

Epithelial (MDA-MB-231)--[EMT]--> mesenchymal (MDA-MB-231)--->invasion/migration/intravasation--{MET}-->epithelial (MDA-MB-231)--->residence and growth in the secondary tumour site

Same goes with any other malignant breast cancer cells. Is this wrong? In respect of aggressiveness, both MCF-7 and MDA-MB231 had the potential to migrate to the lungs, that why they have been isolated from that part. This means they both had the same potential to under go EMT, right?

And if during EMT, will we have a TRUE mesenchymal type of cell (not mesenchymal-like)? or is this "mesenchymal" phrase actually means a mesenchymal behaviour/mesenchymal-like cell?

Weinberg describe transitions between epithelial and mesenchymal states as follow: epithelial -->  epithelial like --> mesenchymal like -->  mesenchymal. For intravasation cell must be in mesenchymal state (www.nature.com/nm/journal/v19/n11/full/nm.3336.html)

MCF 7 is epithelial, but MDAMB 231 is mesenchymal state in vitro model of breast cancer.

P.S. Sorry for my english

Dear All,

I would like to know MDA-MB-436 cell line is mesenchymal or epithelial cell lines???

Hi Myo, and future readers, please check on ATCC website to get information about cell lines. it is safer than asking people.  https://www.atcc.org/Products/All/HTB-130.aspx

it is described as a pleomorphic so you may expect heteregenous phenotype. to confirm you need to, in addition to observe the morphology under microscope, use antibody markers or qPCR of specific gene markers for both epithelial and mesenchymal subtypes.