Table 1.
Summary of metabolic profiles of BxPC-3 (light shaded columns 3–6) and MIA PaCa-2 (dark shaded columns 7–10) pancreatic adenocarcinoma cells (PDAC)
| Metabolite (source-data-matrix-file-log) | Isotopomer fragment dimension | Control | MET | CHS | CHS + MET | Control | MET | CHS | CHS + MET |
|---|---|---|---|---|---|---|---|---|---|
| Glucose content (media-CAS: 50-99-7; 6) | (mg %; mg/100 mL) | 241.33 (±6.75) | 254.67 (±3.33) | 242.33 (±3.25) | 247.67 (±5.01) | 247.33 (±6.37) | 248.00 (±4.77) | 236.67 (±4.04) | 240.67 (±3.79) |
| Glucose consumption (media-CAS: 50-99-7; 7) | (mg %/hour/million cells) | 208.67 (±6.75) | 195.33 (±3.33) | 207.67 (±3.25) | 202.33 (±5.01) | 202.67 (±6.37) | 202.00 (±4.77) | 213.33 (±4.04) | 209.33 (±3.79) |
| Glucose tracer (media-CAS: 138079-87-5; 280) | 13C-labeled fraction (m/z242) (Σm) | 23.47 (±0.04) | 23.64* (±0.12) | 27.87*±(0.04) | 28.02** (±0.0002) | 30.38a (±0.04) | 30.31 (±0.02) | 39.32**, a (±0.005) | 39.26* (±0.004) |
| Glucose tracer (media-CAS: 138079-87-5; 283) | 13C-m2 (m/z242) (m2/Σm) | 97.80(±0.03) | 97.17* (±0.20) | 97.49* (±0.12) | 97.11**(±0.004) | 97.24a (±0.07) | 97.14 (±0.04) | 96.94**, a (±0.07) | 96.93* (±0.12) |
| Lactate (media-CAS: 50-21-5; 20) | 13C-m2 (m/z328) (m2/Σm) | 79.58 (±3.01) | 79.86 (±3.06) | 81.71 ±(3.15) | 77.58 (±3.26) | 91.49 (±4.27) | 83.64 (±3.37) | 80.08 (±3.05) | 84.14 (±3.65) |
| Lactate (media-CAS: 50-21-5; 22B) | Peak-area (abundance × 102) | 4538 (±220) | 3174 (±432) | 1412** ±(78) | 1334** (± 38) | 290a ±(20) | 2055** (±72) | 1827* (±167) | 3456* (± 467) |
| Glutamate (media-CAS: 617-65-2; 78) | 13C-m1 (m/z198) (m1/Σm) | 70.67 (±1.3) | 71.77 (±1.2) | 68.52 (±1.9) | 68.34 (±2.1) | 47.06a (±0.38) | 46.23a (±0.25) | 45.64a (±0.75) | 43.03*, a (±1.05) |
| Glutamate (media-CAS: 617-65-2; 79) | 13C-m2 (m/z198) (m2/Σm) | 28.65 (±1.15) | 27.08 (±0.91) | 30.74 (±1.88) | 31.17 (±2.17) | 38.68a (±1.57) | 43.47a (±1.19) | 43.57a (±1.78) | 46.11*, a (±1.19) |
| Glutamate (media-CAS: 617-65-2; 81) | 13C-m4 (m/z198) (m4/Σm) | 0.29 (±0.03) | 0.25 (±0.02) | 0.27 (±0.02) | 0.32 (±0.01) | 10.14a (±0.25) | 9.65a (±0.55) | 7.62**, a (±0.27) | 6.31**, a (±0.12) |
| Glutamate (media-CAS: 617-65-2; 87B) | Peak-area (abundance) | 150735 (±13440) | 173398 (±16925) | 168611 (±11332) | 87179*,a (±11045) | 25566a (±3095) | 22591a (±3965) | 15879a (±4953) | 15861a (±2906) |
|
Palmitate (pellet-CAS: 57-10-3; 98) |
13C-labeled fraction (m/z270) (Σm) | 8.95 (±0.22) | 8.98 (±0.71) | 8.17 (±0.38) | 3.86** (±0.22) | 4.6a (±0.19) | 5.4a (±0.38) | 15.3**, a (±0.48) | 11.3**, a (±0.41) |
| Palmitate (pellet-CAS: 57-10-3; 101) | Chain elongation-13C-m2 (m/z270) (m2/Σm) | 29.78 (±1.60) | 29.85 (±0.93) | 31.09 (±0.62) | 33.27 (±0.30) | 39.39a (±0.73) | 40.53a (±0.46) | 42.41a (±0.98) | 40.99a (±0.78) |
| Palmitate (pellet-CAS: 57-10-3; 102) | Fraction of new synthesis (FNS) (% of total) | 6.23 (±0.04) | 6.83 (±0.39) | 4.61** (±0.16) | 4.94 (±0.39) | 6.16 (±0.19) | 6.73 (±0.23) | 17.16**, a (±0.57) | 12.31**, a (±0.61) |
| Palmitate (pellet-CAS: 57-10-3; 103) | Ace-CoA enrichment (percent of total) | 11.43 (±0.32) | 11.06 (±0.36) | 11.59 (±1.06) | 5.11** (±0.30) | 5.47a (±0.28) | 6.35a (±0.22) | 9.54** (±0.24) | 9.43**, a (±0.32) |
| Cholesterol (pellet-CAS: 57-88-5; 235) | 13C labeled fraction (Σm) | 17.90 (±0.49) | 17.96 (±1.39) | 0.06** (±0.003) | 0.13** (±0.01) | 9.21a (±0.40) | 10.80a (±0.76) | 0.03**, a (±0.001) | 0.04**, a (±0.002) |
| Cholesterol (pellet-CAS: 57-88-5; 236) | 13C content (Σmn) | 0.57 (±0.06) | 0.57 (±0.09) | 0.02* (±0.001) | 0.01* (±0.0002) | 0.23a (±0.03) | 0.27 (±0.01) | 0.03*, a (± 0.004) | 0.02*, a (±0.002) |
| Cholesterol (pellet- CAS: 57-88-5; 238H) | Peak-area_CHOL(C:27) (abundance × 104) | 3.88 (±0.42) | 3.76 (±0.29) | 7.32** (±0.30) | 7.47* (±0.51) | 2.49 (±0.21) | 2.42a (±0.27) | 5.14*, a (±0.36) | 5.32**, a (±0.28) |
The metabolic profiles of BxPC-3 and MIA PaCa-2 cells in response to 100 μM metformin after 24 h of culture with and without CHS pretreatment for 2 weeks were obtained via SiDMAP analysis using [1,2-13C2]-d-glucose tracer and are shown as Ave ± SD
Source-data-matrix-file-log: source of metabolite, i.e.: culture media or pellets with raw data locator file number
Mn/Σm: Isotopomer/13C labeled fraction as SUM(m 1 + m 2 + .. + m n). Σm n:Molar Enrichment (ME) 13C content as SUM(1 × m 1 + 2 × m 2 + .. + n × m n) (Lee et al. 1992). Number of observations per group: n = 3 (± SD)
CAS Chemical abstracts service registry number
* P < 0.05 versus control
** P < 0.01 versus control
a P < 0.05 versus BxPC-3 (treatment matching comparison between cell lines; where cell lines have been cultured in different media as described in Sect. Methods)