FTIR imaging of MCF-7 colonies and their vicinity in Matrigel-embedded 3D cultures

Abstract

BACKGROUND:

It is now recognized that tumor cells can actively alter their microenvironment and that this remodelled microenvironment can subsequently play a critical role in cancer progression and influence therapeutic responses. To date, the molecular heterogeneity within a 3D cancer cell colony and its influence on the extracellular matrix have not been studied by infrared imaging.

OBJECTIVE:

The objective of this study is to investigate by mid-infrared imaging 3D Matrigel-embedded colonies of pure MCF-7 human mammary adenocarcinoma cell line and the surrounding microenvironment after undergoing formalin fixation and paraffin embedding (FFPE).

METHODS:

In order to better reproduce the procedure used for preservation and storage of clinical tissue specimens for pathologic analysis, 7- and 10-day MCF-7 colonies embedded and grown in Matrigel were FFPE-treated; 4-µm-thick sections were cut, mounted on barium fluoride window and deparaffinized. The Fourier transform infrared (FTIR) images of 4096 spectra were collected in transmission mode using a FPA-based FTIR imaging system. They were pre-processed and analysed by principal component and K-mean cluster analyses.

RESULTS:

At 1654 cm⁻¹ and 1234 cm⁻¹, the intensity of absorption in the colonies is significantly higher than in Matrigel. It can be also noted, on the one hand, that there is a spectral heterogeneity in the intracolonial distribution of the absorbances at 1654, 1644, 1640 and 1634 cm⁻¹ (Amide I range) possibly due to changes in protein secondary structures. On the other hand, we observe that Matrigel close to MCF-7 colonies appears altered with respect to more distant Matrigel matrix.

CONCLUSIONS:

FTIR imaging allowed us to highlight changes in the chemical content in MCF-7 colonies and their direct vicinity in Matrigel-embedded 3D cultures.

Keywords

FTIR imaging 3D cell culture extracellular matrix

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References

[1] Baker

M.J.

Trevisan

Bassan

Bhargava

Butler

H.J.

et al., Using Fourier transform IR spectroscopy to analyze biological materials, Nat. Protoc.9 (2014), 1771–1791.

[2] Bambery

K.R.

Wood

B.R.

and McNaughton

, Resonant Mie scattering (RMieS) correction applied to FTIR images of biological tissue samples, Analyst137 (2012), 126–132.

[3] Barcellos-Hoff

M.H.

and Medina

, New highlights on stroma-epithelial interactions in breast cancer, Breast Cancer Res. Treat.7 (2005), 33–36.

[4] Bassan

Byrne

H.J.

Bonnier

Lee

Dumas

et al., Resonant Mie scattering in infrared spectroscopy of biological materials – Understanding the “dispersion artefact”, Analyst134 (2009), 1586–1593.

[5] Bassan

Kohler

Martens

Lee

Byrne

H.J.

et al., Resonant Mie scattering (RMieS) correction of infrared spectra from highly scattering biological samples, Analyst135 (2010), 268–277.

[6] Bassan

Kohler

Martens

Lee

Jackson

et al., RMieS-EMSC correction for infrared spectra of biological cells: Extension using full Mie theory and GPU computing, J. Biophotonics3 (2010), 609–620.

[7] Bassan

Sachdeva

Kohler

Hughes

Henderson

et al., FTIR microscopy of biological cells and tissue: Data analysis using resonant Mie scattering (RMieS) EMSC algorithm, Analyst137 (2012), 1370–1377.

[8] Benard

Desmedt

Smolina

Szternfeld

Verdonck

et al., Infrared imaging in breast cancer: Automated tissue component recognition and spectral characterization of breast cancer cells as well as the tumor microenvironment, Analyst139 (2014), 1044–1056.

[9] Bhowmick

N.A.

Neilson

E.G.

and Moses

H.L.

, Stromal fibroblasts in cancer initiation and progression, Nature432 (2004), 332–337.

10.

[10] Bird

Miljkovic

and Diem

, Two step resonant Mie scattering correction of infrared micro-spectral data: Human lymph node tissue, J. Biophotonics3 (2010), 597–608.

11.

[11] Cairns

R.A.

Harris

I.S.

and Mak

T.W.

, Regulation of cancer cell metabolism, Nat. Rev. Cancer11 (2011), 85–95.

12.

[12] Che

Z.M.

Jung

T.H.

Choi

J.H.

Yoon

D.J.

Jeong

H.J.

et al., Collagen-based co-culture for invasive study on cancer cells-fibroblasts interaction, Biochem. Biophys. Res. Commun.346 (2006), 268–275.

13.

[13] Erler

J.T.

and Weaver

V.M.

, Three-dimensional context regulation of metastasis, Clin. Exp. Metastasis26 (2009), 35–49.

14.

[14] Gaigneaux

Ruysschaert

J.M.

and Goormaghtigh

, Cell discrimination by attenuated total reflection-Fourier transform infrared spectroscopy: The impact of preprocessing of spectra, Appl. Spectrosc.60 (2006), 1022–1028.

15.

[15] Garamszegi

Garamszegi

S.P.

Shehadeh

L.A.

and Scully

S.P.

, Extracellular matrix-induced gene expression in human breast cancer cells, Mol. Cancer Res.7 (2009), 319–329.

16.

[16] Hanahan

and Weinberg

R.A.

, Hallmarks of cancer: The next generation, Cell144 (2011), 646–674.

17.

[17] Holton

S.E.

Bergamaschi

Katzenellenbogen

B.S.

and Bhargava

, Integration of molecular profiling and chemical imaging to elucidate fibroblast-microenvironment impact on cancer cell phenotype and endocrine resistance in breast cancer, PLoS ONE9 (2014), e96878.

18.

[18] Holton

S.E.

Walsh

M.J.

Kajdacsy-Balla

and Bhargava

, Label-free characterization of cancer-activated fibroblasts using infrared spectroscopic imaging, Biophys. J.101 (2011), 1513–1521.

19.

[19] Kee

N.L.A.

Krause

Blatch

G.L.

Muramoto

Sakka

et al., The proteolytic profile of human cancer procoagulant suggests that it promotes cancer metastasis at the level of activation rather than degradation, Protein J.34 (2015), 338–348.

20.

[20] Khamis

Z.I.

Sahab

Z.J.

Byers

S.W.

and Sang

Q.-X.A.

, Novel stromal biomarkers in human breast cancer tissues provide evidence for the more malignant phenotype of estrogen receptor-negative tumors, J. Biomed. Biotechnol.2011 (2011), 723650.

21.

[21] Khamis

Z.I.

Sahab

Z.J.

and Sang

Q.-X.A.

, Active roles of tumor stroma in breast cancer metastasis, Int. J. Breast Cancer2012 (2012), 574025.

22.

[22] Kleinman

H.K.

and Martin

G.R.

, Matrigel: Basement membrane matrix with biological activity, Semin. Cancer Biol.15 (2005), 378–386.

23.

[23] Kumar

Desmedt

Larsimont

Sotiriou

and Goormaghtigh

, Change in the microenvironment of breast cancer studied by FTIR imaging, Analyst138 (2013), 4058–4065.

24.

[24] Kumar

Shabi

T.S.

and Goormaghtigh

, A FTIR imaging characterization of fibroblasts stimulated by various breast cancer cell lines, PLoS ONE9 (2014), e111137.

25.

[25] Lee

G.Y.

Kenny

P.A.

Lee

E.H.

and Bissell

M.J.

, Three-dimensional culture models of normal and malignant breast epithelial cells, Nat. Methods4 (2007), 359–365.

26.

[26] Lühr

Friedl

Overath

Tholey

Kunze

et al., Mammary fibroblasts regulate morphogenesis of normal and tumorigenic breast epithelial cells by mechanical and paracrine signals, Cancer Lett.325 (2012), 175–188.

27.

[27] Mattson

E.C.

Aboualizadeh

Barabas

M.E.

Stucky

C.L.

and Hirschmugl

C.J.

, Opportunities for live cell FT-infrared imaging: Macromolecule identification with 2D and 3D localization, Int. J. Mol. Sci.14 (2013), 22753–22781.

28.

[28] Olsen

C.J.

Moreira

Lukanidin

E.M.

and Ambartsumian

N.S.

, Human mammary fibroblasts stimulate invasion of breast cancer cells in a three-dimensional culture and increase stroma development in mouse xenografts, BMC Cancer10 (2010), 444.

29.

[29] Poincloux

Lizárraga

and Chavrier

, Matrix invasion by tumour cells: A focus on MT1-MMP trafficking to invadopodia, J. Cell Sci.122 (2009), 3015–3024.

30.

[30] Reddy

L.A.

Mikesh

Moskulak

Harvey

Sherman

et al., Host response to human breast Invasive Ductal Carcinoma (IDC) as observed by changes in the stromal proteome, J. Proteome Res.13 (2014), 4739–4751.

31.

[31] Sadlonova

Mukherjee

Bowe

D.B.

Gault

S.R.

Dumas

N.A.

et al., Human breast fibroblasts inhibit growth of the MCF10AT xenograft model of proliferative breast disease, Am. J. Pathol.170 (2007), 1064–1076.

32.

[32] Schreiber

R.D.

Old

L.J.

and Smyth

M.J.

, Cancer immunoediting: Integrating immunity’s roles in cancer suppression and promotion, Science331 (2011), 1565–1570.

33.

[33] Smolina

and Goormaghtigh

, Infrared imaging of MDA-MB-231 breast cancer cell line phenotypes in 2D and 3D cultures, Analyst140 (2015), 2336–2343.

34.

[34] Studebaker

A.W.

Storci

Werbeck

J.L.

Sansone

Sasser

A.K.

et al., Fibroblasts isolated from common sites of breast cancer metastasis enhance cancer cell growth rates and invasiveness in an interleukin-6-dependent manner, Cancer Res.68 (2008), 9087–9095.

35.

[35] Sung

K.E.

Berthier

Pehlke

Friedl

et al., Understanding the impact of 2D and 3D fibroblast cultures on in vitro breast cancer models, PLoS ONE8 (2013), e76373.

36.

[36] Ueno

Utsumi

Toi

and Shimizu

, Characteristic gene expression profiles of human fibroblasts and breast cancer cells in a newly developed bilateral coculture system, BioMed Res. Int.2015 (2015), 960840.

37.

[37] Wald

Bordry

Foukas

P.G.

Speiser

D.E.

and Goormaghtigh

, Identification of melanoma cells and lymphocyte subpopulations in lymph node metastases by FTIR imaging histopathology, Biochim. Biophys. Acta1862 (2016), 202–212.

38.

[38] Wald

Legat

Meyer

Speiser

D.E.

and Goormaghtigh

, An infrared spectral signature of human lymphocyte subpopulations from peripheral blood, Analyst140 (2015), 2257–2265.

39.

[39] Zaman

M.H.

Trapani

L.M.

Sieminski

A.L.

Siemeski

Mackellar

et al., Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis, Proc. Natl. Acad. Sci. USA103 (2006), 10889–10894.