Infrared imaging of MDA-MB-231 breast cancer cell line phenotypes in 2D and 3D cultures

Smolina, Margarita; Goormaghtigh, Erik

doi:10.1039/c4an01833h

Cited by 21 publications

(12 citation statements)

References 60 publications

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“…In theory, spot density could be increased but their size should remain significantly above the resolution of infrared imaging. As pointed out by earlier [17], IR images are characterized by a poor resolution (with respect to the achievement of visible light microscopy) as expected from the diffraction-limited resolution, e.g. λ ≈ 6 µm at 1654 cm −1 .…”

Section: Discussionmentioning

confidence: 99%

Analysis of protein microarrays by FTIR imaging

Meutter

Derfoufi

Goormaghtigh

2016

BSI

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Abstract. BACKGROUND:Proteins are sensitive to environmental conditions. Whether they are produced for therapeutic purposes or for fundamental research, the integrity of their structure and post-traductional modifications are key issues. Measuring glycosylation or phosphorylation level as well as their secondary structure most often rely on complex and indirect experiments. Infrared spectroscopy presents a series of advantages related to its multivariate character. There is a lack of high-throughput methods able to analyse these parameters. OBJECTIVE: In this paper we attempted to combine protein microarrays and infrared imaging for high throughput analysis of proteins. METHODS: A protein microarrayer was used to produce protein microarrays on BaF 2 slides transparent in the mid-infrared. Spot density was about 25 spots/mm 2 . A 128 × 128 focal plane array infrared detector was used to record images of the protein microarrays. RESULTS: We show that 100 µm diameter spot are easily analyzed. Spots obtained with low protein concentrations, resulting in an average of a single protein monolayer (ca 3 fg/µm 2 for a 66 kDa protein) provided good quality spectra. CONCLUSIONS: Infrared imaging is a label free, high throughput method, able to analyse protein microarrays and to take advantage from the wide information available in the infrared spectra.

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Section: Discussionmentioning

confidence: 99%

Analysis of protein microarrays by FTIR imaging

Meutter

Derfoufi

Goormaghtigh

2016

BSI

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“…Our goal was to find IR patterns that discriminate ECM3 from non-ECM3 tumors ( Fig. 1), as molecular characteristics at the transcriptional level can be related to IR spectra 14,15 . Clinico-pathological characteristics of tumors analyzed are shown in Table 1.…”

Section: Classification Of Ecm3 and Non-ecm3 Tumors By Ft-ir Previoumentioning

confidence: 99%

Infrared Spectroscopic Imaging Visualizes a Prognostic Extracellular Matrix-Related Signature in Breast Cancer

Tiwari

Triulzi

Holton

et al. 2020

Sci Rep

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Molecular analysis techniques such as gene expression analysis and proteomics have contributed greatly to our understanding of cancer heterogeneity. In prior studies, gene expression analysis was shown to stratify patient outcome on the basis of tumor-microenvironment associated genes. A specific gene expression profile, referred to as ECM3 (Extracellular Matrix Cluster 3), indicated poorer survival in patients with grade III tumors. In this work, we aimed to visualize the downstream effects of this gene expression profile onto the tissue, thus providing a spatial context to altered gene expression profiles. Using infrared spectroscopic imaging, we identified spectral patterns specific to the ECM3 gene expression profile, achieving a high spectral classification performance of 0.87 as measured by the area under the curve of the receiver operating characteristic curve. On a patient level, we correctly identified 20 out of 22 ECM3 group patients and 19 out of 20 non-ECM3 group patients by using this spectroscopic imaging-based classifier. By comparing pixels that were identified as ECM3 or non-ECM3 with H&E and IHC images, we were also able to observe an association between tissue morphology and the gene expression clusters, showing the ability of our method to capture broad outcome associated features from infrared images. Breast cancer is the most prevalent cancer in women 1 and is estimated to cause over 40,000 deaths annually in the United States. Although cancer has been thought as a consequence of genetic mutations in an aberrant tissue mass, tumor progression is increasingly viewed as functionally interconnected with the surrounding microenvironment 2. Indeed, crosstalk between cancer cells and the tumor microenvironment components [immune cells, stromal cells, and the extracellular matrix (ECM)] plays a major role in regulating several biological processes 3. Differences in ECM-associated gene expression can describe the biological and clinical heterogeneity of breast cancer, especially in terms of prognosis and response to therapy 2. Recent works have identified a subgroup of breast cancers characterized by overexpression of a robust cluster of genes mainly encoding structural ECM proteins, originally referred to as the ECM3 group 4,5. These features indicate an epithelial to mesenchymal transition (EMT) but with accelerated metastatic potential only in the undifferentiated (grade III) phenotype 4,5. Notably, ECM3 classification provides additional information to assess tumor progression 5. It also identifies tumors in which the interplay between myeloid-derived suppressor cells (MDSCs) and the ECM drives the induction of EMT in tumor cells, and suggests a strategy for inhibition of MDSC by aminobisphosphonates that could revert EMT and lead to less aggressive tumor phenotype 6. Even though such molecular classification is of great importance in stratifying patients, it requires multi-step tissue processing and still requires the validation of an algorithm for classification at the single tumor level to be

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“…Bir titreşim spektroskopisi olan Fourier dönüşümlü kızılötesi (FTIR) spektroskopisi moleküllerin fonksiyonel grupları ile molekül içi/moleküller arası bağlar hakkında doğrudan bilgi sağlamaktadır. IR spektroskopi doku, hücre ve diğer biyolojik örneklerin araştırmalarında yaygın olarak kullanılan biyofiziksel tekniklerden biridir (8)(9)(10)(11)(12)(13)(14)(15)(16). Bu çalışmamızda, MCF-7 meme kanser hücre hattındaki CD44+/CD24-yüzey belirteç özelliği gösteren MKKH"lerin akış sitometrisi ile izole edildikten sonra FTIR spektroskopisi ile hücre biyokimyasındaki farklılaşmalarının ortaya çıkarılması ve moleküler mekanizmanın aydınlatılması amaçlanmıştır.…”

Section: Uluslararasıunclassified

Investigation of breast cancer stem cells in vitro by using IR spectroscopy

et al. 2020

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Öz Amaç: Kanser kök hücreleri (KKH), tümör içinde kendi kendilerini yenileme ve diğer hücre tiplerine farklılaşabilme kapasitesi sebebiyle tümörün başlaması, ilerlemesi, nüksetmesi, metastaz ve terapötik dirence yol açmaktadır. Bu nedenle, meme kanser kök hücrelerinin (MKKH) karakteristik özelliklerinin belirlenmesi gerekmektedir. Bu çalışmanın amacı, MKKH"lerin akış sitometrisi ile izole edildikten sonra Fourier dönüşümlü kızılötesi (FTIR) spektroskopisi kullanarak hücre biyokimyasındaki farklılaşmalarının moleküler seviyede araştırılmasıdır. Gereç ve Yöntem: MCF-7 meme kanser hücre hattındaki CD44+/CD24-yüzey belirteç özelliği gösteren MKKH"ler akış sitometrisi ile izole edilmiştir. MCF10A, MCF-7 kanser hücre (KH) hattı ve bu hattan izole edilen CD44+/CD24-yüzey belirteç özelliklerine sahip MKKH"'ler %0,9 NaCI içerisine resuspanse edildikten sonra FTIR spektrometre ile ölçülmüştür. Bulgular: MCF-7 içerisindeki CD44+/CD24-yüzey belirteç özelliğine sahip KKH"lerinin sort oranı %2,0-2,3 olarak belirlenmiştir. Elde edilen FTIR spektrumlarında, MKKH, meme kanser hücreleri (KH, non-KKH, bulk populasyon) ve sağlıklı hücreler arasında spektral benzerlikler ve farklılıklar tespit edilmiştir. MKKH"lerde lipit ve protein sinyalleri daha güçlü olup hücre zarı akışkanlığı ve dinamiği fazladır. Sağlıklı hücreler ile kıyaslandığında, KH"lerde α-helikal proteinler ve DNA sinyallerinde azalmaya karşın negatif yüklü karboksil gruplarından kaynaklanan sinyallerde artış gözlenmektedir. Bu veriler, MKKH"lerin, sağlıklı ve KH"lere kıyasla yapı, içerik ve dinamiği bakımından oldukça farklı bir profil sergilediğini göstermektedir. Sonuç: Bu çalışma, MKKH"lerinin moleküler yapısı ve içeriğindeki değişikliklerin incelemesi vasıtasıyla terapötik hedefli ilaç çalışmaları yapılabileceğini ortaya koymaktadır. FTIR spektroskopisi boyar madde gerektirmeden, hassas ve hızlı ölçüm alınması, örnek hazırlamada kolaylık ve az miktarda örnek gerektirmesi sebebiyle ileri hücre çalışmalarında ve medikal alanda biyolojik örneklerin analizlerinde kullanılabileceği de gösterilmiştir. Anahtar Sözcükler: Meme kanseri, kanser kök hücresi, FTIR spektroskopisi, hücre zarı akışkanlığı.

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Infrared imaging of MDA-MB-231 breast cancer cell line phenotypes in 2D and 3D cultures

Cited by 21 publications

References 60 publications

Analysis of protein microarrays by FTIR imaging

Analysis of protein microarrays by FTIR imaging

Infrared Spectroscopic Imaging Visualizes a Prognostic Extracellular Matrix-Related Signature in Breast Cancer

Investigation of breast cancer stem cells in vitro by using IR spectroscopy

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