UPLC-Q-TOF/MS-Based Metabolomics Approach Reveals Osthole Intervention in Breast Cancer 4T1 Cells
Abstract
:1. Introduction
2. Results
2.1. Effect of OST on Breast Cancer 4T1 Cell Activity
2.2. Effect of OST on Cell Cycle of Breast Cancer 4T1 Cells
2.3. Effect of OST on Apoptosis of Breast Cancer 4T1 Cells
2.4. Metabolomics Analysis
2.4.1. Quality Control Sample Detection and System Stability Analysis
2.4.2. Metabolite Profiling of 4T1 Cells after OST Treatment
2.5. Multivariate Statistical Analysis and Screening to Identify Potential Marker Metabolites
2.6. Metabolic Pathway Analysis
2.7. Effect of OST on mTOR/SREBP1/FASN Pathway Proteins in Breast Cancer 4T1 Cells
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cell Culture
4.3. Cell Viability Assay
4.4. Cell Cycle Assay
4.5. Cell Apoptosis Assay
4.6. Sample Preparation for Metabolomics
4.7. UPLC-Q-TOF/MS Conditions
4.8. Metabolomics Data Analysis
4.9. Western Blot Analysis
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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NO | M/Z | Compound Name | Chemical Formula | p-Value | Trend |
---|---|---|---|---|---|
1 | 245.1170 | Osthole | C15H16O3 | 0.0001 | ↑ |
2 | 521.3430 | Linoleic acid | C18H32O2 | 0.0048 | ↓ |
3 | 466.3285 | Glycocholic acid | C26H43NO6 | 0.0061 | ↓ |
4 | 468.3065 | PC (14:0/0:0) | C22H46NO7P | 0.0151 | ↓ |
5 | 496.3388 | LysoPC (16:0/0:0) | C24H50NO7P | 0.0151 | ↓ |
6 | 318.2995 | Phytosphingosine | C18H39NO3 | 0.0195 | ─ |
7 | 550.3854 | 1-O-(cis-9-octadecenyl)-2-O-acetyl-sn-glycero-3-phosphocholine | C28H57NO7P+ | 0.0203 | ↓ |
8 | 522.3544 | LysoPC (18:1(9Z)/0:0) | C26H52NO7P | 0.0246 | ↓ |
9 | 521.3430 | Oleic acid | C18H34O2 | 0.0254 | ↓ |
10 | 283.0186 | D-fructose-1-phosphate | C6H13O9P | 0.0361 | ─ |
11 | 204.1222 | L-acetylcarnitine | C9H18NO4 | 0.0407 | ↓ |
12 | 772.6059 | PC (P-18:0/18:1(9Z)) | C44H86NO7P | 0.0445 | ↑ |
13 | 452.2755 | LysoPE (16:0/0:0) | C21H44NO7P | 0.0005 | ↓ |
14 | 303.2319 | N-acetylaspartylglutamate (NAAG) | C11H16N2O8 | 0.0011 | ↓ |
15 | 151.0257 | Oxypurinol | C5H4N4O2 | 0.0017 | ─ |
16 | 154.0617 | His | C6H9N3O2 | 0.0049 | ─ |
17 | 306.0765 | Glutathione | C10H17N3O6S | 0.0071 | ─ |
18 | 195.0508 | Galactonic acid | C6H12O7 | 0.0083 | ─ |
19 | 180.0660 | 3-amino-3-(4-hydroxyphenyl) propionic acid | C9H11NO3 | 0.0092 | ↓ |
20 | 154.0617 | L-histidine | C6H9N3O2 | 0.0098 | ─ |
21 | 155.0097 | Orotic acid | C5H4N2O4 | 0.0156 | ↓ |
22 | 347.1826 | Inosine 5′-monophosphate (IMP) | C10H13N4O8P | 0.0173 | ─ |
23 | 236.0922 | N-lactoyl-phenylalanine | C12H15NO4 | 0.0210 | ─ |
24 | 147.0295 | 3-hydroxyglutaric acid | C5H8O5 | 0.0236 | ↓ |
25 | 509.2864 | 1-oleoyl-2-hydroxy-sn-glycero-3-PG (sodium salt) | C24H47NaO9P | 0.0254 | ─ |
26 | 480.3070 | LysoPE (18:0/0:0) | C23H48NO7P | 0.0298 | ─ |
27 | 535.0338 | UDP-xylose | C14H22N2O16P2 | 0.0302 | ─ |
28 | 171.0260 | Benzylphosphonic acid | C7H9O3P | 0.0315 | ↓ |
29 | 209.0300 | Mucic acid | C6H10O8 | 0.0372 | ↓ |
30 | 402.9918 | Uridine 5′-diphosphate | C9H14N2O12P2 | 0.0390 | ─ |
31 | 771.5151 | Phosphatidylglyceride18:2–18:2 | C42H75O10P | 0.0459 | ↓ |
32 | 167.0204 | Uric acid | C5H4N4O3 | 0.0491 | ─ |
NO | Pathway Name | Total | Expected | Hits | Raw p |
---|---|---|---|---|---|
1 | Phospholipid Biosynthesis | 29 | 0.368 | 2 | 0.0498 |
2 | Methyl Histidine Metabolism | 4 | 0.051 | 1 | 0.0499 |
3 | Pyrimidine Metabolism | 59 | 0.749 | 2 | 0.169 |
4 | Beta Oxidation of Very Long Chain Fatty Acids | 17 | 0.216 | 1 | 0.197 |
5 | Alpha Linolenic Acid and Linoleic Acid Metabolism | 19 | 0.241 | 1 | 0.217 |
6 | Nucleotide Sugar Metabolism | 20 | 0.254 | 1 | 0.227 |
7 | Lactose Synthesis | 20 | 0.254 | 1 | 0.227 |
8 | Purine Metabolism | 74 | 0.939 | 2 | 0.241 |
9 | Androstenedione Metabolism | 24 | 0.305 | 1 | 0.267 |
10 | Estrone Metabolism | 24 | 0.305 | 1 | 0.267 |
11 | Oxidation of Branched Chain Fatty Acids | 26 | 0.330 | 1 | 0.286 |
12 | Starch and Sucrose Metabolism | 31 | 0.394 | 1 | 0.331 |
13 | Ammonia Recycling | 32 | 0.406 | 1 | 0.340 |
14 | Fructose and Mannose Degradation | 32 | 0.406 | 1 | 0.340 |
15 | Amino Sugar Metabolism | 33 | 0.419 | 1 | 0.348 |
16 | Androgen and Estrogen Metabolism | 33 | 0.419 | 1 | 0.348 |
17 | Beta-Alanine Metabolism | 34 | 0.432 | 1 | 0.357 |
18 | Aspartate Metabolism | 35 | 0.444 | 1 | 0.365 |
19 | Galactose Metabolism | 38 | 0.482 | 1 | 0.390 |
20 | Porphyrin Metabolism | 40 | 0.508 | 1 | 0.406 |
21 | Sphingolipid Metabolism | 40 | 0.508 | 1 | 0.406 |
22 | Histidine Metabolism | 43 | 0.546 | 1 | 0.429 |
23 | Bile Acid Biosynthesis | 65 | 0.825 | 1 | 0.576 |
24 | Arachidonic Acid Metabolism | 69 | 0.876 | 1 | 0.598 |
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Li, X.; Zhang, C.; Wu, E.; Han, L.; Deng, X.; Shi, Z. UPLC-Q-TOF/MS-Based Metabolomics Approach Reveals Osthole Intervention in Breast Cancer 4T1 Cells. Int. J. Mol. Sci. 2023, 24, 1168. https://doi.org/10.3390/ijms24021168
Li X, Zhang C, Wu E, Han L, Deng X, Shi Z. UPLC-Q-TOF/MS-Based Metabolomics Approach Reveals Osthole Intervention in Breast Cancer 4T1 Cells. International Journal of Molecular Sciences. 2023; 24(2):1168. https://doi.org/10.3390/ijms24021168
Chicago/Turabian StyleLi, Xiuyun, Chenglun Zhang, Enhui Wu, Liang Han, Xiangliang Deng, and Zhongfeng Shi. 2023. "UPLC-Q-TOF/MS-Based Metabolomics Approach Reveals Osthole Intervention in Breast Cancer 4T1 Cells" International Journal of Molecular Sciences 24, no. 2: 1168. https://doi.org/10.3390/ijms24021168