Comparison of the Metabolite Profiles of HT-29 Colorectal Cancer Cells Treated with Curcumin, Cisplatin, 5-Fluorouracil and Doxorubicin in a Metabolomic Approach
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Abstract
Chemotherapy for colorectal cancer often leads to significant adverse effects on patients, underscoring the need for alternative treatments. Herbal medicines like curcumin are considered a valuable complementary therapy due to their low toxicity profile and potential to mitigate the side effects of chemotherapy. Curcumin's mechanism of action targets multiple pathways, with untargeted metabolomic analysis helping to understand its exact mechanisms and subsequent treatment response. The aim of this study was to compare HT-29 cancer cell metabolites after curcumin and chemotherapy drug interventions to identify metabolites that can predict similar mechanisms of action between these treatments. Principal Component Analysis (PCA) of Fourier transform infrared spectroscopy (FTIR) absorption spectrum showed similar metabolite profiles in HT-29 cell culture media treated with curcumin and the chemotherapeutic cisplatin. Five cell metabolomes emerged after additional gas chromatography mass spectrometry/mass spectrometry (GC-MS/MS) and MS-DIAL data annotation: 1-bromo-2-chloroethane, 2-cyanoacetamide, dimethylamine (DMA), 2-nitrobenzo acid, and butane. The confusion matrix of these five annotated metabolites could be distinguished in HT-29 cell cultures treated with curcumin, but not in control cell cultures or those treated with the drugs cisplatin, doxorubicin, or 5-fluorouracil (5-FU). 2-cyanoacetamide in particular can be used as a marker of HT-29 cells' response to treatment with curcumin based on a p-value of < 0.05. According to these findings, no metabolite can predict the resemblance of curcumin's mechanism of action to chemotherapeutic medicines. Further study should therefore focus on in vivo experimental validation and upgrading metabolomic analysis technologies to further establish the similarities in the metabolite profiles of curcumin and cisplatin treatments.
Keywords: Curcumin, Colorectal Cancer,
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References
1. Effendi-Ys R. Cancer Stem Cells and Molecular Biology Test in Colorectal Cancer: Therapeutic Implications. Vol. 49, Acta Med Indones-Indones J Intern Med. 2017;49(4):351-359.
2. Lin T, Liang C, Peng W, Qiu Y, Peng L. Mechanisms of core Chinese herbs against colorectal cancer: A study based on data mining and network pharmacology.J Evid Based Complementary Altern Med. 2020;8325076:1-15.
3. Bhutadiya Vl, Mistry Kn. A Review On Bioactive Phytochemicals And It’s Mechanism On Cancer Treatment And Prevention By Targeting Multiple Cellular Signaling Pathways. Int J Pharm Pharm Sci. 2021;13(12):1–19.
4. Li P, Pu S, Lin C, He L, Zhao H, Yang C, Guo Z, Xu S, Zhou Z . Curcumin selectively induces colon cancer cell apoptosis and S cell cycle arrest by regulates Rb/E2F/p53 pathway. J Mol Struct. 2022;1263:1-9.
5. Arya N, Prakash OM, Verma AK, Pant AK. Variation In Antioxidant Potential Of Curcuma Longa L. Collected From Different Ecological Niches Of Western Himalayan Region.Int. J. Pharm. Pharm. Sci. 2015; 7(7):1-6.
6. War AR. Curcumin Co-Treatment Sensitizes Multi-Drug Resistant HT29 Colon Cancer Cell Line. J. Can. Res. Immuno-onc. 2018;4(2):1-8.
7. Liu J, Li L, Zhang B, Xu ZP. MnO2-shelled Doxorubicin/Curcumin nanoformulation for enhanced colorectal cancer chemo-immunotherapy. J Colloid Interface Sci. 2022;617:315–325.
8. Woo JH, Park JM, Jang JH, Yang H, Surh YJ, Na HK. Curcumin induces expression of 15-hydroxyprostaglandin dehydrogenase in gastric mucosal cells and mouse stomach in vivo: AP-1 as a potential target. J. Nutr. Biochem. 2020;85:1-11.
9. Krishnamurthy G, Roy D, Kumar J. Curcumin, a natural golden drug and its anticancer aspects from synthesis to delivery: A review. Int. J. Appl. Pharm. Innovare Academics Sciences Pvt. Ltd; 2020; 12:70–84.
10. Huang Y, Liu Z, Liu S, Song F, Jin Y. Studies on the mechanism of Panax Ginseng in the treatment of deficiency of vital energy dementia rats based on urine metabolomics. J Chromatogr B Analyt Technol Biomed Life Sci. 2022;1191:1-9.
11. Gupta M. Synergistic Anti-Cancer Effects Of Natural Products And Their Mode Of Action. Asian j. pharm. clin. res.2021;14:1-7.
12. Altharawi A, Rahman M, Chan KLA. Towards identifying the mode of action of drugs using live-cell FTIR spectroscopy. Analyst. 2019;144(8):2725–2735.
13. Na J, Hwang HJ, Shin MS, Kang M, Lee J, Bang G, Kim YJ, Hwang YJ, Hwang KA, Park YJ. Extract of radish (R. Sativus Linn) promotes anti-atherosclerotic effect using urine metabolomics in ApoE−/− mice. J Funct Foods. 2021;78:1-11.
14. AJDA. Orange data mining Orange with Spectroscopy Add-on Workshop. [Online]. 2018 [cited 2022 Nov 25]. Available from: https://orangedatamining.com/blog/orange-with-spectroscopy-add-on-workshop/.
15. Agarwal A, Kasinathan A, Ganesan R, Balasubramanian A, Bhaskaran J, Suresh S, Srinivasan R, Aravind KB, Sivalingan N. Curcumin induces apoptosis and cell cycle arrest via the activation of reactive oxygen species–independent mitochondrial apoptotic pathway in Smad4 and p53 mutated colon adenocarcinoma HT29 cells. Nutrition Research. 2018;51:67–81.
16. He G, Yang S bing, Wang Y zhong. An integrated chemical characterization based on FT-NIR, and GC–MS for the comparative metabolite profiling of 3 species of the genus Amomum. Anal Chim Acta. 2023;1280:341869.
17. Okafor CE, Ijoma IK, Igboamalu CA, Ezebalu CE, Eze CF, Osita-Chikeze JC, Uzor CE, Ekwuekwe AL. Secondary metabolites, spectra characterization, and antioxidant correlation analysis of the polar and nonpolar extracts of Bryophyllum pinnatum (Lam) Oken. Biotechnologia. 2024;105(2):121–136.
18. Zheng C, Li J, Liu H, Wang Y. Integrating infrared spectroscopy, metabolomics, and network pharmacology to investigate differential metabolites of volatile compounds in porcini mushrooms. J. Food Comp. Anal.2025;139:107164.
19. Tsugawa H, Cajka T, Kind T, Ma Y, Higgins B, Ikeda K, Kanazawa M, Vandergheynst J, Fiehn O, Arita M. MS-DIAL: Data-independent MS/MS deconvolution for comprehensive metabolome analysis. Nat Methods. 2015;12(6):523–526.
20. Jiang L, Sullivan H, Wang B. Principal Component Analysis (PCA) Loading and Statistical Tests for Nuclear Magnetic Resonance (NMR) Metabolomics Involving Multiple Study Groups. Anal Lett. 2022;55(10):1648–1662.
21. Martinelli DD. Machine learning for metabolomics research in drug discovery. Intelligence-Based Medicine Elsevier B.2023;8:100101 .
22. UCSC. WPMUCDN Nuclear Magnetic Resonance (NMR). [Online]. 2015 [cited 2022 Nov 25]. Available from: https://cpb-use1.wpmucdn.com/sites.ucsc.edu/dist/9/291/files/2015/11/108M-Lecture-3-NMR-Intro-Tables-2.pdf
23. Wu BB, Gong YP, Wu XH, Chen YY, Chen FF, Jin LT, Cheng BR, Hu F, Xiong B. Fourier transform infrared spectroscopy for the distinction of MCF-7 cells treated with different concentrations of 5-fluorouracil. J Transl Med. 2015;13(108):1-8.
24. Li G, Fang S, Shao X, Li Y, Tong Q, Kong B, Chen L, Wang Y, Yang J, Yu H, Xie X, Zhang J. Curcumin reverses NNMT-induced 5-fluorouracil resistance via increasing ROS and cell cycle arrest in colorectal cancer cells. Biomolecules. 2021;11(9):1295.
25. Zhang F, Zhang Y, Zhao W, Deng K, Wang Z, Yang C, Ma L, Openkova MS, Hou Y, Li K. Metabolomics for biomarker discovery in the diagnosis, prognosis, survival and recurrence of colorectal cancer: a systematic review. Oncotarget. 2017;8(21):35460–35472.
26. Genua F, Mirković B, Mullee A, Levy M, Gallagher WM, Vodicka P, Hughes DJ. Association of circulating short chain fatty acid levels with colorectal adenomas and colorectal cancer. Clin Nutr ESPEN. 2021;46:297–304.
27. Debik J, Sangermani M, Wang F, Madssen TS, Giskeødegård GF. Multivariate analysis of NMR‐based metabolomic data. NMR Biomed. 2021:1-21.
28. Kellog JJ, Graf TN, Paine MF, McCune JS, Kvalheim OM, Oberlies NH, Cech NB. Comparison of Metabolomics Approaches for Evaluating the Variability of Complex Botanical Preparations: Green Tea (Camellia sinensis) as a Case Study. J Nat Prod. 2017; 80(5):1457-1466.
29. Tran TQ, Hanse EA, Habowski AN, Li H, Ishak Gabra MB, Yang Y, Lowman Xh, Ooi Am, Liao SY, Edwards RA, Waterman ML, Kong M. α-Ketoglutarate attenuates Wnt signaling and drives differentiation in colorectal cancer. Nat Cancer. 2020;1(3):345–358.
30. Farhadi P, Yarani R, Dokaneheifard S, Mansouri K. The emerging role of targeting cancer metabolism for cancer therapy. J. Tum. Bio. 2020;42(10):1-18.
31. Hammad A, Elshaer M, Tang X. Identification of potential biomarkers with colorectal cancer based on bioinformatics analysis and machine learning. Math Biosci Eng. 2021;18(6):8997–9015.
32. Schrimpe-Rutledge AC, Codreanu SG, Sherrod SD, McLean JA. Untargeted Metabolomics Strategies—Challenges and Emerging Directions. J Am Soc Mass Spectrom. 2016;27(12):1897–1905.
33. Arasaradnam RP, Mcfarlane MJ, Ryan-Fisher C, Westenbrink E, Hodges P, Thomas MG, Chambers S, Oconnell N, Bailey C, Harmston C, Nwokolo CU, Bardhan KD, Covington JA. Detection of colorectal cancer (CRC) by urinary volatile organic compound analysis. PLoS One. 2014;9(9):1-6.
34. Bednarz-Misa I, Fleszar MG, Zawadzki M, Kapturkiewicz B, Kubiak A, Neubauer K, Witkiewicz W, Korpacka-Krzystek M. L-arginine/no pathway metabolites in colorectal cancer: Relevance as disease biomarkers and predictors of adverse clinical outcomes following surgery. J Clin Med. 2020;9(6):1–32.
35. Tsikas D. Urinary dimethylamine (DMA) and its precursor asymmetric dimethylarginine (ADMA) in clinical medicine, in the context of nitric oxide (no) and beyond. J. Clin. Med. 2020;9(6):1843.
36. Zhang N, Gao M, Wang Z, Zhang J, Cui W, Li J, Zhu X, Zhang H, Yang DH, Xu X. Curcumin reverses doxorubicin resistance in colon cancer cells at the metabolic level. J Pharm Biomed Anal. 2021;15:201.
37. LeCun Y, Bengio Y, Hinton G. Deep learning. Nature. 2015;521(7553):436–44.
38. Bradley M. ThermoFisher FTIR Basic Organic Functional Group Reference Chart. [Online]. 2015 [cited 2023 Feb 9]. Available from: https://www.thermofisher.com/blog/materials/a-gift-for-you-an-ftir-basic-organic-functional-group-reference-chart/