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In situ characterizing membrane lipid phenotype of breast cancer cells using mass spectrometry profiling.

He M, Guo S, Li Z - Sci Rep (2015)

Bottom Line: Lipid composition in cell membrane is closely associated with cell characteristics.The Western blotting assays indicated that the expression of five lipogenesis-related enzymes (i.e., fatty acid synthase 1(FASN1), stearoyl-CoA desaturase 1 (SCD1), stearoyl-CoA desaturase 5 (SCD5), choline kinase α (CKα), and sphingomyelin synthase 1) was associated with the types of the breast cells, and that the SCD1 level in MCF-7 cells was significantly increased relative to other breast cell lines.Our findings suggest that elevated expression levels of FASN1, SCD1, SCD5, and CKα may closely correlated with enhanced levels of saturated and monounsaturated lipids in breast cancer cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences &School of Basic Medicine, Peking Union Medical College, Beijing 100005, P.R. China.

ABSTRACT
Lipid composition in cell membrane is closely associated with cell characteristics. Here, matrix-assisted laser desorption/ionization- Fourier transform ion cyclotron resonance mass spectrometry was employed to in situ determine membrane components of human mammary epithelial cells (MCF-10 A) and six different breast cancer cell lines (i.e., BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-MB-157, and MDA-MB-361) without any lipid extraction and separation. Partial least-square discriminant analysis indicated that changes in the levels of these membrane lipids were closely correlated with the types of breast cell lines. Elevated levels of polyunsaturated lipids in MCF-10 A cells relative to six breast cancer cells and in BT-20 cells relative to other breast cancer cell lines were detected. The Western blotting assays indicated that the expression of five lipogenesis-related enzymes (i.e., fatty acid synthase 1(FASN1), stearoyl-CoA desaturase 1 (SCD1), stearoyl-CoA desaturase 5 (SCD5), choline kinase α (CKα), and sphingomyelin synthase 1) was associated with the types of the breast cells, and that the SCD1 level in MCF-7 cells was significantly increased relative to other breast cell lines. Our findings suggest that elevated expression levels of FASN1, SCD1, SCD5, and CKα may closely correlated with enhanced levels of saturated and monounsaturated lipids in breast cancer cell lines.

No MeSH data available.


Related in: MedlinePlus

PLS-DA score plots of membrane lipids detected in this study between MCF-10A cells and breast cancer cell lines.(A) PLS-DA model analysis of 180 common metabolites to differentiate six different breast cancer cell lines (i.e., BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-BR-157, and MDA-BR-361) from MCF-10A control. (B) Significantly changed levels of 8 common lipids (i.e., PC(32:1), SM(34:0), C22:4, PC(38:4), PI(38:4), PC(30:0), C18:2, and C16:1) with the VIP values of >1.0 between MCF-10A and six different breast cancer cell lines. (C) PLS-DA model analysis of the 180 common metabolites from six different breast cancer cell lines to differentiate six breast cancer cell lines. (D) Significantly changed levels of 15 common lipids (i.e., PE(38:4), PE(P-38:5), PE(36:4), PE(O-38:5), C22:6, PI(34:1), PI(36:1), PC(36:1), C20:4, PC(36:2), PC(28:0), PC(34:1), SM(34:1), PE(36:2), and PC(32:0)) with the VIP values of >1.0 between six breast cancer cell lines.
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f2: PLS-DA score plots of membrane lipids detected in this study between MCF-10A cells and breast cancer cell lines.(A) PLS-DA model analysis of 180 common metabolites to differentiate six different breast cancer cell lines (i.e., BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-BR-157, and MDA-BR-361) from MCF-10A control. (B) Significantly changed levels of 8 common lipids (i.e., PC(32:1), SM(34:0), C22:4, PC(38:4), PI(38:4), PC(30:0), C18:2, and C16:1) with the VIP values of >1.0 between MCF-10A and six different breast cancer cell lines. (C) PLS-DA model analysis of the 180 common metabolites from six different breast cancer cell lines to differentiate six breast cancer cell lines. (D) Significantly changed levels of 15 common lipids (i.e., PE(38:4), PE(P-38:5), PE(36:4), PE(O-38:5), C22:6, PI(34:1), PI(36:1), PC(36:1), C20:4, PC(36:2), PC(28:0), PC(34:1), SM(34:1), PE(36:2), and PC(32:0)) with the VIP values of >1.0 between six breast cancer cell lines.

Mentions: In this study, an untargeted membrane lipid profiling was performed to in situ analyze intact cell samples using MALDI-FTICR MS platform. Representative mass spectra of MCF-10 A cells are shown in Fig. 1. Finally, a total of 180 membrane lipids including 69 detected in the positive ion mode and 111 detected in the negative ion mode were observed in seven different breast cell lines, and the detailed information on their m/z values are listed in Supplementary Table S1. PLS-DA score plot revealed the obvious differences between control (MCF-10A) and six different breast cancer cell lines (BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-MB-157, and MDA-MB-361), with the predicted residual sum of square (PRESS) of 0.1697 (Fig. 2A), and 46 lipids have the variable importance in the projection (VIP) values of >1.0 (Supplementary Table S2). It should be noted that 8 simultaneously up- or down-regulated lipids in breast cancer cells relative to control were observed, and they are PC(32:1), SM(34:0), C22:4, PC(38:4), PI(38:4), PC(30:0), C18:2, and C16:1. The detailed information on their identification is shown in Supplementary Fig. S1 and Table S3. As shown in Fig. 2B, the levels of PC(32:1), PC(30:0), and C16:1 in breast cancer cells were significantly up-regulated, while the levels of SM(34:0), PC(38:4), PI(38:4), C18:2, and C22:4 in breast cancer cells were remarkably down-regulated relative to control. Their detailed change trends are shown in Supplementary Fig. S2A.


In situ characterizing membrane lipid phenotype of breast cancer cells using mass spectrometry profiling.

He M, Guo S, Li Z - Sci Rep (2015)

PLS-DA score plots of membrane lipids detected in this study between MCF-10A cells and breast cancer cell lines.(A) PLS-DA model analysis of 180 common metabolites to differentiate six different breast cancer cell lines (i.e., BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-BR-157, and MDA-BR-361) from MCF-10A control. (B) Significantly changed levels of 8 common lipids (i.e., PC(32:1), SM(34:0), C22:4, PC(38:4), PI(38:4), PC(30:0), C18:2, and C16:1) with the VIP values of >1.0 between MCF-10A and six different breast cancer cell lines. (C) PLS-DA model analysis of the 180 common metabolites from six different breast cancer cell lines to differentiate six breast cancer cell lines. (D) Significantly changed levels of 15 common lipids (i.e., PE(38:4), PE(P-38:5), PE(36:4), PE(O-38:5), C22:6, PI(34:1), PI(36:1), PC(36:1), C20:4, PC(36:2), PC(28:0), PC(34:1), SM(34:1), PE(36:2), and PC(32:0)) with the VIP values of >1.0 between six breast cancer cell lines.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4462148&req=5

f2: PLS-DA score plots of membrane lipids detected in this study between MCF-10A cells and breast cancer cell lines.(A) PLS-DA model analysis of 180 common metabolites to differentiate six different breast cancer cell lines (i.e., BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-BR-157, and MDA-BR-361) from MCF-10A control. (B) Significantly changed levels of 8 common lipids (i.e., PC(32:1), SM(34:0), C22:4, PC(38:4), PI(38:4), PC(30:0), C18:2, and C16:1) with the VIP values of >1.0 between MCF-10A and six different breast cancer cell lines. (C) PLS-DA model analysis of the 180 common metabolites from six different breast cancer cell lines to differentiate six breast cancer cell lines. (D) Significantly changed levels of 15 common lipids (i.e., PE(38:4), PE(P-38:5), PE(36:4), PE(O-38:5), C22:6, PI(34:1), PI(36:1), PC(36:1), C20:4, PC(36:2), PC(28:0), PC(34:1), SM(34:1), PE(36:2), and PC(32:0)) with the VIP values of >1.0 between six breast cancer cell lines.
Mentions: In this study, an untargeted membrane lipid profiling was performed to in situ analyze intact cell samples using MALDI-FTICR MS platform. Representative mass spectra of MCF-10 A cells are shown in Fig. 1. Finally, a total of 180 membrane lipids including 69 detected in the positive ion mode and 111 detected in the negative ion mode were observed in seven different breast cell lines, and the detailed information on their m/z values are listed in Supplementary Table S1. PLS-DA score plot revealed the obvious differences between control (MCF-10A) and six different breast cancer cell lines (BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-MB-157, and MDA-MB-361), with the predicted residual sum of square (PRESS) of 0.1697 (Fig. 2A), and 46 lipids have the variable importance in the projection (VIP) values of >1.0 (Supplementary Table S2). It should be noted that 8 simultaneously up- or down-regulated lipids in breast cancer cells relative to control were observed, and they are PC(32:1), SM(34:0), C22:4, PC(38:4), PI(38:4), PC(30:0), C18:2, and C16:1. The detailed information on their identification is shown in Supplementary Fig. S1 and Table S3. As shown in Fig. 2B, the levels of PC(32:1), PC(30:0), and C16:1 in breast cancer cells were significantly up-regulated, while the levels of SM(34:0), PC(38:4), PI(38:4), C18:2, and C22:4 in breast cancer cells were remarkably down-regulated relative to control. Their detailed change trends are shown in Supplementary Fig. S2A.

Bottom Line: Lipid composition in cell membrane is closely associated with cell characteristics.The Western blotting assays indicated that the expression of five lipogenesis-related enzymes (i.e., fatty acid synthase 1(FASN1), stearoyl-CoA desaturase 1 (SCD1), stearoyl-CoA desaturase 5 (SCD5), choline kinase α (CKα), and sphingomyelin synthase 1) was associated with the types of the breast cells, and that the SCD1 level in MCF-7 cells was significantly increased relative to other breast cell lines.Our findings suggest that elevated expression levels of FASN1, SCD1, SCD5, and CKα may closely correlated with enhanced levels of saturated and monounsaturated lipids in breast cancer cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences &School of Basic Medicine, Peking Union Medical College, Beijing 100005, P.R. China.

ABSTRACT
Lipid composition in cell membrane is closely associated with cell characteristics. Here, matrix-assisted laser desorption/ionization- Fourier transform ion cyclotron resonance mass spectrometry was employed to in situ determine membrane components of human mammary epithelial cells (MCF-10 A) and six different breast cancer cell lines (i.e., BT-20, MCF-7, SK-BR-3, MDA-MB-231, MDA-MB-157, and MDA-MB-361) without any lipid extraction and separation. Partial least-square discriminant analysis indicated that changes in the levels of these membrane lipids were closely correlated with the types of breast cell lines. Elevated levels of polyunsaturated lipids in MCF-10 A cells relative to six breast cancer cells and in BT-20 cells relative to other breast cancer cell lines were detected. The Western blotting assays indicated that the expression of five lipogenesis-related enzymes (i.e., fatty acid synthase 1(FASN1), stearoyl-CoA desaturase 1 (SCD1), stearoyl-CoA desaturase 5 (SCD5), choline kinase α (CKα), and sphingomyelin synthase 1) was associated with the types of the breast cells, and that the SCD1 level in MCF-7 cells was significantly increased relative to other breast cell lines. Our findings suggest that elevated expression levels of FASN1, SCD1, SCD5, and CKα may closely correlated with enhanced levels of saturated and monounsaturated lipids in breast cancer cell lines.

No MeSH data available.


Related in: MedlinePlus