Dandelion root extracts and taraxasterol inhibit LPS‑induced colorectal cancer cell viability by blocking TLR4‑NF&kappa;B‑driven ACE2 and TMPRSS2 pathways

Yang,Kerry; Wang,Yuehong

doi:10.3892/etm.2024.12544

Dandelion root extracts and taraxasterol inhibit LPS‑induced colorectal cancer cell viability by blocking TLR4‑NFκB‑driven ACE2 and TMPRSS2 pathways

Authors:
- Kerry Yang
- Yuehong Wang
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Affiliations: Faculty of Health Sciences, McMaster University, Hamilton, Ontario L8S 4L8, Canada, Cardiovascular and Metabolic Research Unit, Laurentian University, Sudbury, Ontario P3E 2C6, Canada
Published online on: April 19, 2024 https://doi.org/10.3892/etm.2024.12544
Article Number: 256
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Colorectal cancer is the fourth leading cause of cancer‑related death worldwide. Notably, abnormalities in intestinal bacteria may contribute to the initiation or progression of colorectal cancer. Lipopolysaccharide (LPS), a bacterial endotoxin, is elevated in patients with colorectal cancer. The present study investigated the protective effects of dandelion root extracts and taraxasterol (TS; a major pharmacologically active compound in dandelion root extracts) on LPS‑induced colorectal cancer cell viability, as well as the underlying mechanisms. Cell viability was assessed by MTT assay, and protein and gene expression levels were determined by western blotting and quantitative PCR. It was revealed that LPS at a low dose (0.5 µg/ml) significantly promoted the viability of human colorectal cancer cells but did not affect normal colon epithelial cells. The addition of dandelion root extracts (0.1‑1 mg/ml) or TS (0.05‑1 µg/ml) was able to reverse the LPS‑induced increase in colorectal cancer cell viability and colony formation. Mechanistically, dandelion root extracts or TS may inhibit the LPS‑promoted toll‑like receptor 4 (TLR4)/NFκB‑p65 pathway and transcription levels of pro‑inflammatory genes (TNFα, IL4 and IL6). Compared with normal colon epithelial cells, human colorectal cancer cells had higher expression levels of angiotensin‑converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), which could be further enhanced by LPS treatment but this was reversed by co‑incubation with dandelion root extracts or TS. In addition, suppression of the TLR4/NFκB‑p65 pathway with CLI095 significantly reversed the stimulatory effect of LPS on the expression levels of ACE2 and TMPRSS2, whereas TNFα (10 ng/ml) markedly induced the expression levels of ACE2 and TMPRSS2. In conclusion, the present study suggested that dandelion root extracts and TS could be used as prevention strategies for reversing bacteria‑driven colorectal cancer cell viability.

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1	Reis SAD, da Conceição LL and Peluzio MDCG: Intestinal microbiota and colorectal cancer: Changes in the intestinal microenvironment and their relation to the disease. J Med Microbiol. 68:1391–1407. 2019.PubMed/NCBI View Article : Google Scholar
2	Maryam S, Krukiewicz K, Haq IU, Khan AA, Yahya G and Cavalu S: Interleukins (cytokines) as biomarkers in colorectal cancer: Progression, detection, and monitoring. J Clin Med. 12(3127)2023.PubMed/NCBI View Article : Google Scholar
3	Hofseth LJ, Hebert JR, Chanda A, Chen H, Love BL, Pena MM, Murphy EA, Sajish M, Sheth A, Buckhaults PJ and Berger FG: Early-onset colorectal cancer: Initial clues and current views. Nat Rev Gastroenterol Hepatol. 17:352–364. 2020.PubMed/NCBI View Article : Google Scholar
4	Collins D, Hogan AM and Winter DC: Microbial and viral pathogens in colorectal cancer. Lancet Oncol. 12:504–512. 2011.PubMed/NCBI View Article : Google Scholar
5	de Waal GM, de Villiers WJS, Forgan T, Roberts T and Pretorius E: Colorectal cancer is associated with increased circulating lipopolysaccharide, inflammation and hypercoagulability. Sci Rep. 10(8777)2020.PubMed/NCBI View Article : Google Scholar
6	Tjalsma H, Boleij A, Marchesi JR and Dutilh BE: A bacterial driver-passenger model for colorectal cancer: Beyond the usual suspects. Nat Rev Microbiol. 10:575–582. 2012.PubMed/NCBI View Article : Google Scholar
7	Lucas C, Barnich N and Nguyen HTT: Microbiota, inflammation and colorectal cancer. Int J Mol Sci. 18(1310)2017.PubMed/NCBI View Article : Google Scholar
8	Chu FF, Esworthy RS, Chu PG, Longmate JA, Huycke MM, Wilczynski S and Doroshow JH: Bacteria-induced intestinal cancer in mice with disrupted Gpx1 and Gpx2 genes. Cancer Res. 64:962–968. 2004.PubMed/NCBI View Article : Google Scholar
9	Pandey H, Tang DWT, Wong SH and Lal D: Gut microbiota in colorectal cancer: Biological role and therapeutic opportunities. Cancers (Basel). 15(866)2023.PubMed/NCBI View Article : Google Scholar
10	Mandal P: Molecular mechanistic pathway of colorectal carcinogenesis associated with intestinal microbiota. Anaerobe. 49:63–70. 2018.PubMed/NCBI View Article : Google Scholar
11	Kasai C, Sugimoto K, Moritani I, Tanaka J, Oya Y, Inoue H, Tameda M, Shiraki K, Ito M, Takei Y and Takase K: Comparison of human gut microbiota in control subjects and patients with colorectal carcinoma in adenoma: Terminal restriction fragment length polymorphism and next-generation sequencing analyses. Oncol Rep. 35:325–333. 2016.PubMed/NCBI View Article : Google Scholar
12	Pitari GM, Zingman LV, Hodgson DM, Alekseev AE, Kazerounian S, Bienengraeber M, Hajnóczky G, Terzic A and Waldman SA: Bacterial enterotoxins are associated with resistance to colon cancer. Proc Natl Acad Sci USA. 100:2695–2699. 2003.PubMed/NCBI View Article : Google Scholar
13	Abdelhamid AM, Deeb ME and Zaafan MA: The protective effect of xanthenone against LPS-induced COVID-19 acute respiratory distress syndrome (ARDS) by modulating the ACE2/Ang-1-7 signaling pathway. Eur Rev Med Pharmacol Sci. 26:5285–5296. 2022.PubMed/NCBI View Article : Google Scholar
14	Seo GS: The role of NF-kappaB in colon cancer. Korean J Gastroenterol. 57:3–7. 2011.PubMed/NCBI View Article : Google Scholar : (In Korean).
15	O'Leary DP, Bhatt L, Woolley JF, Gough DR, Wang JH, Cotter TG and Redmond HP: TLR-4 signalling accelerates colon cancer cell adhesion via NF-κB mediated transcriptional up-regulation of Nox-1. PLoS One. 7(e44176)2012.PubMed/NCBI View Article : Google Scholar
16	Killeen SD, Wang JH, Andrews EJ and Redmond HP: Bacterial endotoxin enhances colorectal cancer cell adhesion and invasion through TLR-4 and NF-kappaB-dependent activation of the urokinase plasminogen activator system. Br J Cancer. 100:1589–1602. 2009.PubMed/NCBI View Article : Google Scholar
17	Ranneh Y, Akim AM, Hamid HA, Khazaai H, Fadel A and Mahmoud AM: Stingless bee honey protects against lipopolysaccharide induced-chronic subclinical systemic inflammation and oxidative stress by modulating Nrf2, NF-κB and p38 MAPK. Nutr Metab (Lond). 16(15)2019.PubMed/NCBI View Article : Google Scholar
18	An X, Lin W, Liu H, Zhong W, Zhang X, Zhu Y, Wang X, Li J and Sheng Q: SARS-CoV-2 host receptor ACE2 protein expression atlas in human gastrointestinal tract. Front Cell Dev Biol. 9(659809)2021.PubMed/NCBI View Article : Google Scholar
19	Bernardi S, Zennaro C, Palmisano S, Velkoska E, Sabato N, Toffoli B, Giacomel G, Buri L, Zanconati F, Bellini G, et al: Characterization and significance of ACE2 and Mas receptor in human colon adenocarcinoma. J Renin Angiotensin Aldosterone Syst. 13:202–209. 2012.PubMed/NCBI View Article : Google Scholar
20	Liu C, Wang K, Zhang M, Hu X, Hu T, Liu Y, Hu Q, Wu S and Yue J: High expression of ACE2 and TMPRSS2 and clinical characteristics of COVID-19 in colorectal cancer patients. NPJ Precis Oncol. 5(1)2021.PubMed/NCBI View Article : Google Scholar
21	Reindl-Schwaighofer R, Eskandary F, Bartko J, Heinzel A, Jilma B, Hecking M and Schoergenhofer C: Corticosteroid treatment prevents lipopolysaccharide-induced increase of ACE2 and reduces fibrin degradation products in bronchoalveolar lavage fluid. Front Med (Lausanne). 9(856891)2022.PubMed/NCBI View Article : Google Scholar
22	Burgueño JF, Reich A, Hazime H, Quintero MA, Fernandez I, Fritsch J, Santander AM, Brito N, Damas OM, Deshpande A, et al: Expression of SARS-CoV-2 entry molecules ACE2 and TMPRSS2 in the gut of patients with IBD. Inflamm Bowel Dis. 26:797–808. 2020.PubMed/NCBI View Article : Google Scholar
23	Zabiegala A, Kim Y and Chang KO: Roles of host proteases in the entry of SARS-CoV-2. Anim Dis. 3(12)2023.PubMed/NCBI View Article : Google Scholar
24	Ahmadi M, Pashangzadeh S, Mousavi P, Saffarzadeh N, Habibi MA, Hajiesmaeili F and Rezaei N: ACE2 correlates with immune infiltrates in colon adenocarcinoma: Implication for COVID-19. Int Immunopharmacol. 95(107568)2021.PubMed/NCBI View Article : Google Scholar
25	Hfaiedh M, Brahmi D and Zourgui L: Hepatoprotective effect of Taraxacum officinale leaf extract on sodium dichromate-induced liver injury in rats. Environ Toxicol. 31:339–349. 2016.PubMed/NCBI View Article : Google Scholar
26	Sigstedt SC, Hooten CJ, Callewaert MC, Jenkins AR, Romero AE, Pullin MJ, Kornienko A, Lowrey TK, Slambrouck SV and Steelant WF: Evaluation of aqueous extracts of Taraxacum officinale on growth and invasion of breast and prostate cancer cells. Int J Oncol. 32:1085–1090. 2008.PubMed/NCBI
27	Ovadje P, Ammar S, Guerrero JA, Arnason JT and Pandey S: Dandelion root extract affects colorectal cancer proliferation and survival through the activation of multiple death signalling pathways. Oncotarget. 7:73080–73100. 2016.PubMed/NCBI View Article : Google Scholar
28	Ding A and Wen X: Dandelion root extract protects NCM460 colonic cells and relieves experimental mouse colitis. J Nat Med. 72:857–866. 2018.PubMed/NCBI View Article : Google Scholar
29	Sharma K and Zafar R: Occurrence of taraxerol and taraxasterol in medicinal plants. Pharmacogn Rev. 9:19–23. 2015.PubMed/NCBI View Article : Google Scholar
30	Liu B, He Z, Wang J, Xin Z, Wang J, Li F and Fu Y: Taraxasterol inhibits LPS-induced inflammatory response in BV2 microglia cells by activating LXRα. Front Pharmacol. 9(278)2018.PubMed/NCBI View Article : Google Scholar
31	Che L, Li Y, Song R, Qin C, Hao W, Wang B, Yang L, Peng P and Xu F: Anti-inflammatory and anti-apoptosis activity of taraxasterol in ulcerative colitis in vitro and in vivo. Exp Ther Med. 18:1745–1751. 2019.PubMed/NCBI View Article : Google Scholar
32	Zhang X, Xiong H, Li H and Cheng Y: Protective effect of taraxasterol against LPS-induced endotoxic shock by modulating inflammatory responses in mice. Immunopharmacol Immunotoxicol. 36:11–16. 2014.PubMed/NCBI View Article : Google Scholar
33	Cho SY, Park JY, Park EM, Choi MS, Lee MK, Jeon SM, Jang MK, Kim MJ and Park YB: Alternation of hepatic antioxidant enzyme activities and lipid profile in streptozotocin-induced diabetic rats by supplementation of dandelion water extract. Clin Chim Acta. 317:109–117. 2002.PubMed/NCBI View Article : Google Scholar
34	Barrow K, Wang Y, Yu R, Zhu J and Yang G: H₂S protects from oxidative stress-driven ACE2 expression and cardiac aging. Mol Cell Biochem. 477:1393–1403. 2022.PubMed/NCBI View Article : Google Scholar
35	Li L, Li M, Li Y, Sun W, Wang Y, Bai S, Li H, Wu B, Yang G, Wang R, et al: Exogenous H₂S contributes to recovery of ischemic post-conditioning-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2-STAT3 pathways in the aging cardiomyocytes. Cell Biosci. 6(26)2016.PubMed/NCBI View Article : Google Scholar
36	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
37	Yang Y, Yang L, Jiang S, Yang T, Lan J, Lei Y, Tan H and Pan K: HMGB1 mediates lipopolysaccharide-induced inflammation via interacting with GPX4 in colon cancer cells. Cancer Cell Int. 20(205)2020.PubMed/NCBI View Article : Google Scholar
38	Yang K and Zhang Y: Reversal of heavy metal-induced antibiotic resistance by dandelion root extracts and taraxasterol. J Med Microbiol. 69:1049–1061. 2020.PubMed/NCBI View Article : Google Scholar
39	Sun Q, Liu Q, Zheng Y and Cao X: Rapamycin suppresses TLR4-triggered IL-6 and PGE(2) production of colon cancer cells by inhibiting TLR4 expression and NF-kappaB activation. Mol Immunol. 45:2929–2936. 2008.PubMed/NCBI View Article : Google Scholar
40	Ying J, Zhou H, Wang Z, You Q, Chen J, Lu H and Zhang J: Aspirin increases chemosensitivity of colorectal cancer cells and inhibits the expression of toll-like receptor 4. Cancer Cell Int. 23(6)2023.PubMed/NCBI View Article : Google Scholar
41	Zhuo Q, Yu B, Zhou J, Zhang J, Zhang R, Xie J, Wang Q and Zhao S: Lysates of Lactobacillus acidophilus combined with CTLA-4-blocking antibodies enhance antitumor immunity in a mouse colon cancer model. Sci Rep. 9(20128)2019.PubMed/NCBI View Article : Google Scholar
42	Li Z, Wang K, Ji X, Wang H and Zhang Y: ACE2 suppresses the inflammatory response in LPS-induced porcine intestinal epithelial cells via regulating the NF-κB and MAPK pathways. Peptides. 149(170717)2022.PubMed/NCBI View Article : Google Scholar
43	Lin X, Lin W, Zhuang Y and Gao F: Angiotensin-converting enzyme 2 inhibits lipopolysaccharide-caused lung fibrosis via downregulating the transforming growth factor β-1/Smad2/Smad3 pathway. J Pharmacol Exp Ther. 381:236–246. 2022.PubMed/NCBI View Article : Google Scholar
44	Cheng J, Zhou J, Fu S, Fu J, Zhou B, Chen H, Fu J and Wei C: Prostate adenocarcinoma and COVID-19: The possible impacts of TMPRSS2 expressions in susceptibility to SARS-CoV-2. J Cell Mol Med. 25:4157–4165. 2021.PubMed/NCBI View Article : Google Scholar
45	Klooster JPT, Bol-Schoenmakers M, van Summeren K, van Vliet ALW, de Haan CAM, van Kuppeveld FJM, Verkoeijen S and Pieters R: Enterocytes, fibroblasts and myeloid cells synergize in anti-bacterial and anti-viral pathways with IL22 as the central cytokine. Commun Biol. 4(631)2021.PubMed/NCBI View Article : Google Scholar
46	Tran HTT, Gigl M, Le NPK, Dawid C and Lamy E: In vitro effect of Taraxacum officinale leaf aqueous extract on the interaction between ACE2 cell surface receptor and SARS-CoV-2 spike protein D614 and four mutants. Pharmaceuticals (Basel). 14(1055)2021.PubMed/NCBI View Article : Google Scholar
47	Zhang X, Jia F, Ma W, Li X and Zhou X: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells. Vet Res. 53(104)2022.PubMed/NCBI View Article : Google Scholar