Enhanced detection of metastatic prostate cancer cells in human plasma with lipid bodies staining

BMC Cancer

Thuc T Le¹  Oscar B Goodman²  Ranjana Mitra² 

[1]Desert Research Institute, 10530 Discovery Drive, Las Vegas, NV 89135, USA

[2]Roseman University of Health Sciences, 11 Sunset Way, Henderson, NV 89014, USA

关键词: Proteomics;    Protein O-linked glycosylation;    Protein lysine acetylation;    Prostate cancer;    Lipid bodies;    Flow cytometry;    Circulating tumor cell;    Coherent anti-Stokes Raman microscopy;    Cancer energy metabolism;   

Others  :  859066
DOI  :  10.1186/1471-2407-14-91

 received in 2013-11-13, accepted in 2014-02-11,  发布年份 2014

【 摘 要 】

Background

Reprogramming of energy metabolism of malignant cancer cells confers competitive advantage in growth environments with limited resources. However, not every process of cancer development is associated with competition for resources. During hematogenous transport, cancer cells are exposed to high levels of oxygen and nutrients. Does energy metabolism of cancer cells change as a function of exposure to the bloodstream? Could such changes be exploited to improve the detection of circulating tumor cells (CTC)? These questions have clinical significance, but have not yet been sufficiently examined.

Methods

The energy metabolism was examined as a function of incubation in nutrient-rich plasma in prostate metastatic cancer cells LNCaP and non-transformed prostate epithelial cells RWPE1. Uptake kinetics of a fluorescent glucose analog (2-NBD) and lipophilic dyes (DiD & Bodipy) were measured in both cell lines, as well as in peripheral blood mononuclear cells (PBMC).

Results

LNCaP cells exhibited hyper-acetylation of low molecular weight proteins compared to RWPE1 cells. Following plasma incubation, protein lysine acetylation profile was unchanged for LNCaP cells while significantly altered for RWPE1 cells. O-linked glycosylated protein profiles were different between LNCaP and RWPE1 cells and varied in both cell lines with plasma incubation. Maximal respiration or glycolytic capacities was unchanged in LNCaP cells and impaired in RWPE1 cells following plasma incubation. However, the uptake rates of 2-NBD and DiD were insufficient for discrimination of LNCaP, or RWPE1 cells from PBMC. On the other hand, both RWPE1 and LNCaP cells exhibited intracellular lipid bodies following plasma incubation; whereas, PBMC did not. The presence of lipid bodies in LNCaP cells permitted retention of Bodipy dye and allowed discrimination of LNCaP cells from PBMC with flow cytometry.

Conclusions

Despite clear differences in energy metabolism, metastatic prostate cancer cells could not be efficiently distinguished from non-transformed prostate epithelial cells using fluorescent glucose or lipid uptake kinetics. However, metastatic prostate cancer cells in plasma could be clearly distinguished from blood nucleated cells due to the presence of intracellular lipid bodies. Fluorescent labeling of lipid bodies permitted a simple and sensitive means for high throughput detection of metastatic prostate cancer cells in human plasma.

【 授权许可】

   
2014 Mitra et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Hanahan D, Weinberg RA: Hallmarks of cancer: the next generation. Cell 2011, 144(5):646-674.
• [2]Hsu PP, Sabatini DM: Cancer cell metabolism: Warburg and beyond. Cell 2008, 134(5):703-707.
• [3]Koppenol WH, Bounds PL, Dang CV: Otto Warburg’s contributions to current concepts of cancer metabolism. Nat Rev Cancer 2011, 11(5):325-337.
• [4]Cairns RA, Harris IS, Mak TW: Regulation of cancer cell metabolism. Nat Rev Cancer 2011, 11(2):85-95.
• [5]Wellen KE, Thompson CB: A two-way street: reciprocal regulation of metabolism and signalling. Nat Rev Mol Cell Biol 2012, 13(4):270-276.
• [6]Vander Heiden MG, Cantley LC, Thompson CB: Understanding the Warburg effect: The metabolic requirements of cell proliferation. Science 2009, 234:1029-1033.
• [7]Semenza GL: Hypoxia-inducible factors in physiology and medicine. Cell 2012, 148(3):399-408.
• [8]Dang CV: Links between metabolism and cancer. Genes Dev 2012, 26(9):877-890.
• [9]Bristow RG, Hill RP: Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability. Nat Rev Cancer 2008, 8(3):180-192.
• [10]Gupta GP, Massague J: Cancer metastasis: building a framework. Cell 2006, 127(4):679-695.
• [11]Pantel K, Brakenhoff RH, Brandt B: Detection, clinical relevance and specific biological properties of disseminating tumour cells. Nat Rev Cancer 2008, 8(5):329-340.
• [12]Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, Miller MC, Reuben JM, Doyle GV, Allard WJ, Terstappen LW, et al.: Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 2004, 351(8):781-791.
• [13]Faltas B: Cornering metastases: therapeutic targeting of circulating tumor cells and stem cells. Front Oncol 2012, 2:68.
• [14]Yu M, Stott S, Toner M, Maheswaran S, Haber DA: Circulating tumor cells: approaches to isolation and characterization. J Cell Biol 2011, 192(3):373-382.
• [15]Zhang J, Nuebel E, Wisidagama DR, Setoguchi K, Hong JS, Van Horn CM, Imam SS, Vergnes L, Malone CS, Koehler CM, et al.: Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells. Nat Protoc 2012, 7(6):1068-1085.
• [16]Urasaki Y, Johlfs MG, Fiscus RR, Le TT: Imaging immune and metabolic cells of visceral adipose tissues with multimodal nonlinear optical microscopy. PLoS One 2012, 7(6):e38418.
• [17]Le TT, Yue S, Cheng JX: Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy. J Lipid Res 2010, 51(11):3091-3102.
• [18]Evans CL, Xie XS: Coherent anti-stokes raman scattering microscopy: chemically selective imaging for biology and medicine. Annu Rev Anal Chem 2008, 1(1):883-909.
• [19]Mitra R, Chao O, Urasaki Y, Goodman OB, Le TT: Detection of lipid-rich prostate circulating tumour cells with coherent anti-Stokes Raman scattering microscopy. BMC Cancer 2012, 12:540. BioMed Central Full Text
• [20]Liu Y, Zuckier LS, Ghesani NV: Dominant uptake of fatty acid over glucose by prostate cells: a potential new diagnostic and therapeutic approach. Anticancer Res 2010, 30(2):369-374.
• [21]Alipour A, van Oostrom AJ, Izraeljan A, Verseyden C, Collins JM, Frayn KN, Plokker TW, Elte JW, Castro Cabezas M: Leukocyte activation by triglyceride-rich lipoproteins. Arterioscler Thromb Vasc Biol 2008, 28(4):792-797.
• [22]Wu H, Gower RM, Wang H, Perrard XY, Ma R, Bullard DC, Burns AR, Paul A, Smith CW, Simon SI, et al.: Functional role of CD11c + monocytes in atherogenesis associated with hypercholesterolemia. Circulation 2009, 119(20):2708-2717.
• [23]Le TT, Huff TB, Cheng JX: Coherent anti-Stokes Raman scattering imaging of lipids in cancer metastasis. BMC Cancer 2009, 9:42. BioMed Central Full Text
• [24]Pezacki JP, Blake JA, Danielson DC, Kennedy DC, Lyn RK, Singaravelu R: Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy. Nat Chem Biol 2011, 7(3):137-145.
• [25]Le TT, Rehrer CW, Huff TB, Nichols MB, Camarillo IG, Cheng JX: Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma. Mol Imaging 2007, 6(3):205-211.
• [26]Chowdary PD, Jiang Z, Chaney EJ, Benalcazar WA, Marks DL, Gruebele M, Boppart SA: Molecular histopathology by spectrally reconstructed nonlinear interferometric vibrational imaging. Cancer Res 2010, 70(23):9562-9569.
• [27]Yang Y, Li F, Gao L, Wang Z, Thrall MJ, Shen SS, Wong KK, Wong ST: Differential diagnosis of breast cancer using quantitative, label-free and molecular vibrational imaging. Biomed Opt Express 2011, 2(8):2160-2174.
• [28]Meyer T, Guntinas-Lichius O, von Eggeling F, Ernst G, Akimov D, Schmitt M, Dietzek B, Popp J: Multimodal nonlinear microscopic investigations on head and neck squamous cell carcinoma: Toward intraoperative imaging. Head Neck 2012, 35(9):E280-287.
• [29]Galli R, Sablinskas V, Dasevicius D, Laurinavicius A, Jankevicius F, Koch E, Steiner G: Non-linear optical microscopy of kidney tumours. J Biophotonics 2013, 7(1-2):23-27.
• [30]Salameh TS, Le TT, Nichols MB, Bauer E, Cheng J, Camarillo IG: An ex vivo co-culture model system to evaluate stromal-epithelial interactions in breast cancer. Int J Cancer 2013, 132(2):288-296.