Supplemental Materials
Metabolic Responses of HeLa Cells to Silica Nanoparticles by NMR-based Metabolomic Analyses
Running title: Metabolic response of HeLa cells to silica nanoparticles
Jianghua Feng1, *, Jinquan Li1, Huifeng Wu2, Zhong Chen1
1Department of Electronic Science, Fujian Provincial Key Laboratory of Plasma and Magnetic Resonance, XiamenUniversity, Xiamen, 361005, China
2Key Laboratory of Coastal Zone Environment Processes, CAS, Shandong Provincial Key Laboratory of Coastal Zone Environment Processes, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, 264003, China
* Correspondence to: Jianghua Feng
Email: , Tel: +86-592-2186393, Fax: +86-592-2186393
R2X=44.5%, Q2=0.584
R2X=55.5%, Q2=0.857
FigureS1.OPLS-DA scores plots (left panel) and corresponding coefficient plots (middle and right panels) derived from the 1H HRMAS CPMG NMR spectra of HeLa cells obtained from different pair-wise groups. The color maps show the significance of metabolite variations between the two classes. Peaks in the positive direction indicate metabolites that are more abundant in the dosed groups than control group. Consequently, metabolites that are more abundant in the control group are presented as peaks in the negative direction. C, control groups; L, low-dosage groups; H, high-dosage groups; 6, 6 h post-treatment; 48, 48 h post-treatment with SiO2 nano-particles. Keys: Ace, acetate; Cit, citrate; Cr, Creatine; Gln, glutamine; Glu, glutamate; Ileu, isoleucine; Lac, lactate; Leu, leucine; Lys, lysine; m-I, myo-inositol; MA, methylamine; Met, methonine; PC, phosphocholine; Phe, phenylalanine; Tyr, tyrosine; Val, valine.
R2X=51.8%, R2Y=99.0%, Q2=0.908
R2X=54.7%, R2Y=98.1%, Q2=0.824
R2X=44.5%, R2Y=97.7%, Q2=0.546
R2X=46.2%, R2Y=98.8%, Q2=0.879
R2X=42.7%, R2Y=98.7%, Q2=0.862
R2X=55.5%, R2Y=99.1%, Q2=0.935
Figure S2.PLS-DA scores plots (left) and corresponding plots (right) of permutation tests (n=100) derived from the 1H HRMAS CPMG NMR spectra of HeLa cells obtained from different pair-wise groups. C, control groups; L, low-dosage groups; H, high-dosage groups; 6, 6 h post-treatment; 48, 48 h post-treatment with SiO2 nano-particles
R2X=75.8%, Q2=0.838
R2X=59.2%, Q2=0.914
FigureS3.OPLS-DA scores plots (left panel) and corresponding coefficient plots (middle and right panels) derived from the 1H NOESY NMR spectra of cell growth media obtained from different pair-wise groups. The color maps show the significance of metabolite variations between the two classes. Peaks in the positive direction indicate metabolites that are more abundant in the dosed groups than control group. Consequently, metabolites that are more abundant in the control group are presented as peaks in the negative direction. C, control groups; L, low-dosage groups; H, high-dosage groups; 6, 6 h post-treatment; 48, 48 h post-treatment with SiO2 nano-particles. Keys: Ace, acetate; Ala, alanine; For, formate; Glc, Glucose; Gln, glutamine; HIB, α-hydrxoyisobutyrate; IB, isobutyrate; Ileu, isoleucine; Lac, lactate; Leu, leucine; Lys, lysine; Met, methonine; PC, phosphocholine; Phe, phenylalanine; Pro, proline; Py, Pyruvate; TMAO, trimethylamine N-oxide; Val, valine.
R2X=84.6%, R2Y=93.2%, Q2=0.869
R2X=63.7%, R2Y=93.3%, Q2=0.715
R2X=75.8%, R2Y=90.8%, Q2=0.755
R2X=58.0%, R2Y=89.3%, Q2=0.506
R2X=63.7%, R2Y=96.3%, Q2=0.823
R2X=59.2%, R2Y=97.8%, Q2=0.917
Figure S4.PLS-DA scores plots (left) and corresponding plots (right) of permutation tests (n=100) derived from the 1H NOESY NMR spectra of cell growth media obtained from different pair-wise groups. C, control groups; L, low-dosage groups; H, high-dosage groups; 6, 6 h post-treatment; 48, 48 h post-treatment with SiO2 nano-particles
Table S1The 1H NMR data of the metabolites in the HeLa cell and growth medium
No. / Metabolites / 1H Shift (multiplicity) / Moiety / Sample1a / Isoleucine / 0.94(tb); 1.01(d); 1.26(m);
1.46(m); 1.98(m); 3.67(d) / δ-CH3; β-CH3; γ-CH;
γ-CH´; β-CH; α-CH / Mc, C
2 / Leucine / 0.96(d); 0.97(d);
1.70(m); 3.75(m) / δ-CH3; δ-CH´3;
CH2&γ-CH; α-CH / M, C
3 / Valine / 0.99(d); 1.04(d);
2.28(m); 3.61(d) / γ-CH3; γ-CH´3;
β-CH; α-CH / M, C
4 / Isobutyrate / 1.13(d); 2.55(m) / CH3; α-CH / M
5 / Lactate / 1.33(d); 4.12(q) / CH3; CH / M, C
6 / Lysine / 1.46(m); 1.73(m);1.91(m);
3.03(t); 3.76(t) / γ-CH2; δ-CH2; β-CH2;
ε-CH2; α-CH / M, C
7 / Alanine / 1.48(d); 3.78(q) / CH3; CH / M, C
8 / Acetate / 1.92(s) / CH3 / M, C
9 / Proline / 2.03(m); 2.35(m);
3.37(m); 4.18(m) / β-CH&γ-CH2; β-CH´;
δ-CH2; α-CH / M
10 / Glutamine / 2.14(m); 2.46(m); 3.78(t) / β-CH2; γ-CH2; α-CH / M, C
11 / Methionine / 2.14(s); 2.16(m);
2.65(t); 3.86(t) / S-CH3; β-CH2;
S-CH2; α-CH / M, C
12 / Acetone / 2.23(s) / CH3 / M
13 / Pyruvate / 2.37(s) / CH3 / M
14 / Succinate / 2.41(s) / CH2 / M, C
15 / Choline / 3.19(s); 3.52(m); 4.07(m) / CH3; N-CH2; O-CH2 / M, C
16 / Phosphocholine / 3.20(s); 3.53(m); 4.14(m) / CH3; N-CH2; O-CH2 / M, C
17 / Glycerophosphocholine / 3.35(s); 3.81(m); 4.30(m) / CH3; N-CH2; P-O-CH2 / M
18 / β-Glucose / 3.25(dd); 3.41(t); 3.46(m);
3.49(t); 3.72(dd); 3.90(dd);
4.65(d) / C-H2; C-H4; C-H5;
C-H3; C-H6; C-H6´;
C-H1 / M
19 / α-Glucose / 3.42(t); 3.54(dd); 3.71(t);
3.74(m); 3.84(m); 5.24(d) / C-H4; C-H2; C-H3;
C-H6; C-H5&6´; C-H1 / M
20 / TMAO / 3.29(s) / CH3 / M
21 / Glycine / 3.56(s) / CH2 / M, C
22 / Threonine / 1.33(d); 3.59(d); 4.25(m) / CH3; α-CH; β-CH / M
23 / Tyrosine / 3.06(m); 3.20(m); 3.94(m); 6.92(d); 7.20(d) / β-CH; β-CH´; α-CH;
m-CH; o-CH / M, C
24 / Phenylalanine / 3.13(m); 3.28(m); 4.00(m); 7.33(d); 7.38(t); 7.43(m) / β-CH; β-CH´; α-CH;
o-CH; p-CH; m-CH / M, C
25 / Histidine / 3.14(m); 3.25(m); 3.99(m); 7.08(s); 7.83(s) / β-CH; β-CH´; α-CH;
CH4; CH2 / M
26 / Tryptophan / 3.31(m); 3.49(m); 4.06(m);
7.03(m); 7.35(s); 7.55(d);
7.60(m); 7.74(d) / β-CH; β-CH´; α-CH;
CH8; CH2; CH6;
CH7; CH9 / M
27 / Formate / 8.46(s) / CH / M
28 / α-Hydroxyisobutyrate / 1.37(s) / CH3 / M
HEPES / 2.82(br); 2.94(m); 2.97(t);
3.03(br); 3.16(m); 3.84(t) / M
Eth / Ethanol / 1.19(t); 3.66(q) / CH3; CH2 / M
U1 / Unassigned 1 / 1.14(d) / M
U2 / Unassigned 2 / 1.58(d) / M
HDL / Cholesterol / 0.76(br) / C
L1 / VLDL&LDL / 0.90(br) / CH3-(CH2)n- / C
L2 / VLDL&LDL / 1.31(br) / CH3-(CH2)n- / C
L3 / Triglyceride / 1.59(br) / -CH2-CH2-C=O / C
U3 / Unassigned 3 / 1.77(br) / C
L4 / Triglyceride / 2.03(br) / -CH2-CH=CH- / C
L5 / Triglyceride / 2.06(br) / -CH2-CH=CH- / C
29 / Glutamate / 2.10(m); 2.35(m); 3.78(t) / β-CH2; γ-CH2; α-CH / C
L6 / Triglyceride / 2.26(br) / -CH2-CH2-C=O / C
30 / Citrate / 2.55(d), 2.69(d) / CH2 / C
31 / Methylamine / 2.56(s) / CH3 / C
L7 / Triglyceride / 2.80(br) / =CH-CH2-CH= / C
32 / Creatine / 3.04(s); 3.93(s) / CH3; CH2 / C
33 / myo-Inositol / 3.25(t); 3.61(dd); 4.07(t) / C
34 / Glycerol / 3.65(m); / C
U4 / Unassigned 4 / 3.91(m) / C
L8 / Triglyceride / 5.33(br) / -CH=CH- / C
U5 / Unassigned 5 / 5.91(m) / C
35 / ADP / 5.99(s); 7.88(d); 8.19(s) / C
36 / ATP / 6.11(s); 8.24(s); 8.35(s) / C
a The numbering system is the same as used in Fig. 1.
b Multiplicity: s, singlet; d, doublet; t, triplet; q, quartet; dd, doublet of doublets; m, multiplet; br, broad resonance.
c M, cell growth medium; C, HeLa cells
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