Raman and coherent anti-Stokes Raman scattering microscopy studies of changes in lipid content and composition in hormone-treated breast and prostate cancer cells

Abstract. Increasing interest in the role of lipids in cancer cell proliferation and resistance to drug therapies has motivated the need to develop better tools for cellular lipid analysis. Quantification of lipids in cells is typically done by destructive chromatography protocols that do not provide spatial information on lipid distribution and prevent dynamic live cell studies. Methods that allow the analysis of lipid content in live cells are therefore of great importance. Using micro-Raman spectroscopy and coherent anti-Stokes Raman scattering (CARS) microscopy, we generated a lipid profile for breast (T47D, MDA-MB-231) and prostate (LNCaP, PC3) cancer cells upon exposure to medroxyprogesterone acetate (MPA) and synthetic androgen R1881. Combining Raman spectra with CARS imaging, we can study the process of hormone-mediated lipogenesis. Our results show that hormone-treated cancer cells T47D and LNCaP have an increased number and size of intracellular lipid droplets and higher degree of saturation than untreated cells. MDA-MB-231 and PC3 cancer cells showed no significant changes upon treatment. Principal component analysis with linear discriminant analysis of the Raman spectra was able to differentiate between cancer cells that were treated with MPA, R1881, and untreated.

[1]  J. Renwick Beattie,et al.  A critical evaluation of Raman spectroscopy for the analysis of lipids: Fatty acid methyl esters , 2004, Lipids.

[2]  Michitaka Suzuki,et al.  Lipid droplets: a classic organelle with new outfits , 2008, Histochemistry and Cell Biology.

[3]  S. Shapiro,et al.  Vasectomy and the risk of prostate cancer. , 1990, American journal of epidemiology.

[4]  D. McLean,et al.  Real-time Raman Spectroscopy for in Vivo Skin Cancer Diagnosis Raman Spectroscopy of Skin Cancer , 2022 .

[5]  M. Zern,et al.  Label‐free imaging and analysis of the effects of lipolysis products on primary hepatocytes , 2011, Journal of biophotonics.

[6]  Wenlung Chen,et al.  Structural Analysis of Triacylglycerols and Edible Oils by Near-Infrared Fourier Transform Raman Spectroscopy , 2003, Applied spectroscopy.

[7]  F. Kuhajda,et al.  Fatty acid synthase and cancer: new application of an old pathway. , 2006, Cancer research.

[8]  Andrew Folick,et al.  Label-free imaging of lipid dynamics using Coherent Anti-stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS) microscopy. , 2011, Current opinion in genetics & development.

[9]  Claudio R. Santos,et al.  Lipid metabolism in cancer , 2012, The FEBS journal.

[10]  M. Cox,et al.  Effect of simvastatin on castration-resistant prostate cancer cells , 2014, Lipids in Health and Disease.

[11]  Ji-Xin Cheng,et al.  Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy , 2010, Journal of Lipid Research.

[12]  À. Sierra,et al.  The Lipid Phenotype of Breast Cancer Cells Characterized by Raman Microspectroscopy: Towards a Stratification of Malignancy , 2012, PloS one.

[13]  Annika Enejder,et al.  Monitoring of lipid storage in Caenorhabditis elegans using coherent anti-Stokes Raman scattering (CARS) microscopy , 2007, Proceedings of the National Academy of Sciences.

[14]  S. Yamaguchi,et al.  Long-chain fatty acids induce lipid droplet formation in a cultured human hepatocyte in a manner dependent of Acyl-CoA synthetase. , 2006, Biological & pharmaceutical bulletin.

[15]  K. Parker,et al.  Practical methods for noise removal: applications to spikes, nonstationary quasi-periodic noise, and baseline drift. , 2009, Analytical chemistry.

[16]  D. Chalbos,et al.  Progestin-induced fatty acid synthetase in human mammary tumors: From molecular to clinical studies , 1992, The Journal of Steroid Biochemistry and Molecular Biology.

[17]  Isabel R Schlaepfer,et al.  Progestin modulates the lipid profile and sensitivity of breast cancer cells to docetaxel , 2012, Molecular and Cellular Endocrinology.

[18]  T. Huser,et al.  Raman spectroscopic analysis of biochemical changes in individual triglyceride-rich lipoproteins in the pre- and postprandial state. , 2005, Analytical chemistry.

[19]  Stephen T. C. Wong,et al.  Label-free high-resolution imaging of prostate glands and cavernous nerves using coherent anti-Stokes Raman scattering microscopy , 2011, Biomedical optics express.

[20]  F. Clavel-Chapelon,et al.  Dietary fat intake and development of specific breast cancer subtypes. , 2014, Journal of the National Cancer Institute.

[21]  J. Swinnen,et al.  Androgens, lipogenesis and prostate cancer , 2004, The Journal of Steroid Biochemistry and Molecular Biology.

[22]  T. T. Le,et al.  Coherent anti-Stokes Raman scattering imaging of lipids in cancer metastasis , 2009, BMC Cancer.

[23]  T. Huser,et al.  Fatty Acids from Very Low-Density Lipoprotein Lipolysis Products Induce Lipid Droplet Accumulation in Human Monocytes , 2010, The Journal of Immunology.

[24]  Ji-Xin Cheng,et al.  Cholesteryl ester accumulation induced by PTEN loss and PI3K/AKT activation underlies human prostate cancer aggressiveness. , 2014, Cell metabolism.

[25]  J. Borén,et al.  Lipid droplets as dynamic organelles connecting storage and efflux of lipids. , 2009, Biochimica et biophysica acta.

[26]  T. B. Bakker Schut,et al.  Discriminating basal cell carcinoma from perilesional skin using high wave-number Raman spectroscopy. , 2007, Journal of biomedical optics.

[27]  Caroline H. Diep,et al.  Progesterone receptor-cyclin D1 complexes induce cell cycle-dependent transcriptional programs in breast cancer cells. , 2014, Molecular endocrinology.

[28]  R. Fisher.  THE USE OF MULTIPLE MEASUREMENTS IN TAXONOMIC PROBLEMS , 1936 .

[29]  John Paul Pezacki,et al.  Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy , 2011, Nature chemical biology.

[30]  Christopher J. Frank,et al.  Raman spectroscopy of normal and diseased human breast tissues. , 1995, Analytical chemistry.

[31]  Andreas Zumbusch,et al.  Coherent anti-Stokes Raman scattering microscopy , 1999 .

[32]  P. Baade,et al.  International epidemiology of prostate cancer: geographical distribution and secular trends. , 2009, Molecular nutrition & food research.

[33]  Sumeet Mahajan,et al.  Coherent anti-Stokes Raman scattering for label-free biomedical imaging , 2013 .

[34]  K. Natter,et al.  Yeast and cancer cells – common principles in lipid metabolism , 2013, Biochimica et biophysica acta.

[35]  Yong Q. Chen,et al.  Lipids and prostate cancer. , 2012, Prostaglandins & other lipid mediators.

[36]  O. Warburg.  [Origin of cancer cells]. , 1956, Oncologia.

[37]  Nektarios Tavernarakis,et al.  Label-Free Imaging of Lipid Depositions in C. elegans Using Third-Harmonic Generation Microscopy , 2014, PloS one.

[38]  H. Abramczyk,et al.  Raman imaging at biological interfaces: applications in breast cancer diagnosis , 2013, Molecular Cancer.