Regulation of the migration of colorectal cancer stem cells via the TLR4/MyD88 signaling pathway by the novel surface marker CD14 following LPS stimulation

Li,Yufei; Shi,Jiayi; Liu,Zhixin; Lin,Yonggang; Xie,An; Sun,Wenxiu; Liu,Jiaqi; Liang,Jun

doi:10.3892/ol.2023.14194

Regulation of the migration of colorectal cancer stem cells via the TLR4/MyD88 signaling pathway by the novel surface marker CD14 following LPS stimulation

Authors:
- Yufei Li
- Jiayi Shi
- Zhixin Liu
- Yonggang Lin
- An Xie
- Wenxiu Sun
- Jiaqi Liu
- Jun Liang
View Affiliations

Affiliations: Morphology Laboratory, Mudanjiang Medical University, Mudanjiang, Heilongjiang 157011, P.R. China, School of Life Sciences, Mudanjiang Medical University, Mudanjiang, Heilongjiang 157011, P.R. China, Department of Extracorporeal Circulation, Mudanjiang Cardiovascular Disease Hospital, Mudanjiang, Heilongjiang 157011, P.R. China
Published online on: December 18, 2023 https://doi.org/10.3892/ol.2023.14194
Article Number: 60
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cell surface markers are most widely used in the study of cancer stem cells (CSCs). However, cell surface markers that are safely and stably expressed in CSCs have yet to be identified. Colonic CSCs express leukocyte CD14. CD14 binding to the ligand lipopolysaccharide (LPS) is involved in the inflammatory response via the Toll‑like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88) signaling pathway. TLR4 and MyD88 have been reported to promote the proliferation, metastasis and tumorigenicity of colon cancer cells, which is consistent with the characteristics of CSCs. In the present study, the proposed experimental method to detect cell proliferation, metastasis and tumorigenesis was used to confirm that, under LPS stimulation, CD14 promoted the proliferation, migration and tumorigenesis of colonic CSCs via the TLR4/MyD88 signaling pathway. Cell Counting Kit‑8 and 5‑ethynyl‑2'‑deoxyuridine assays were used to assess the proliferation and migration of the cells. Colony formation and nude mouse xenograft assays were used to assess the capacity of cells to form tumors. Using western blotting and reverse transcription‑quantitative PCR, the mRNA and protein levels of CD14, TLR4 and MyD88 were examined. It was confirmed that CD14 promoted the proliferation, metastasis and tumorigenesis of colon CSCs in response to LPS stimulation via the TLR4/MyD88 signaling pathway, and CD14⁺ colon cancer cells were successfully isolated and sorted. According to the results of proliferation assay, it was determined that CD14 regulated the LPS‑induced proliferation of colon CSCs. CD14, TLR4 and MyD88 protein and mRNA expression was upregulated in colon CSCs in response to LPS stimulation. This indicates a potential novel target for colon CSC‑related studies.

View Figures

View References

1	Ahmad R, Singh JK, Wunnava A, Al-Obeed O, Abdulla M and Srivastava SK: Emerging trends in colorectal cancer: Dysregulated signaling pathways (Review). Int J Mol Med. 47:142021. View Article : Google Scholar : PubMed/NCBI
2	Lewandowska A, Rudzki G, Lewandowski T, Stryjkowska-Góra A and Rudzki S: Title: risk factors for the diagnosis of colorectal cancer. Cancer Control. 29:107327482110566922022. View Article : Google Scholar : PubMed/NCBI
3	Salibasic M, Pusina S, Bicakcic E, Pasic A, Gavric I, Kulovic E, Rovcanin A and Beslija S: Colorectal cancer surgical treatment, our experience. Med Arch. 73:412–414. 2019. View Article : Google Scholar : PubMed/NCBI
4	Li Y, Wu M, Xu S, Huang H, Yan L and Gu Y: Colorectal cancer stem cell-derived exosomal long intergenic noncoding RNA 01315 (LINC01315) promotes proliferation, migration, and stemness of colorectal cancer cells. Bioengineered. 13:10827–10842. 2022. View Article : Google Scholar : PubMed/NCBI
5	Batlle E and Clevers H: Cancer stem cells revisited. Nat Med. 23:1124–1134. 2017. View Article : Google Scholar : PubMed/NCBI
6	Hervieu C, Christou N, Battu S and Mathonnet M: The role of cancer stem cells in colorectal cancer: From the basics to novel clinical trials. Cancers (Basel). 13:10922021. View Article : Google Scholar : PubMed/NCBI
7	Kopenhaver J, Crutcher M, Waldman SA and Snook AE: The shifting paradigm of colorectal cancer treatment: A look into emerging cancer stem cell-directed therapeutics to lead the charge toward complete remission. Expert Opin Biol Ther. 21:1335–1345. 2021. View Article : Google Scholar : PubMed/NCBI
8	Gupta R, Bhatt LK, Johnston TP and Prabhavalkar KS: Colon cancer stem cells: Potential target for the treatment of colorectal cancer. Cancer Biol Ther. 20:1068–1082. 2019. View Article : Google Scholar : PubMed/NCBI
9	Munro MJ, Wickremesekera SK, Peng L, Tan ST and Itinteang T: Cancer stem cells in colorectal cancer: A review. J Clin Pathol. 71:110–116. 2018. View Article : Google Scholar : PubMed/NCBI
10	Yu X, Lin Y, Yan X, Tian Q, Li L and Lin EH: CD133, stem cells, and cancer stem cells: Myth or Reality? Curr Colorectal Cancer Rep. 7:253–259. 2011. View Article : Google Scholar : PubMed/NCBI
11	Janova H, Böttcher C, Holtman IR, Regen T, van Rossum D, Götz A, Ernst AS, Fritsche C, Gertig U, Saiepour N, et al: CD14 is a key organizer of microglial responses to CNS infection and injury. Glia. 64:635–649. 2016. View Article : Google Scholar : PubMed/NCBI
12	Ciesielska A, Matyjek M and Kwiatkowska K: TLR4 and CD14 trafficking and its influence on LPS-induced pro-inflammatory signaling. Cell Mol Life Sci. 78:1233–1261. 2021. View Article : Google Scholar : PubMed/NCBI
13	Hsu RY, Chan CH, Spicer JD, Rousseau MC, Giannias B, Rousseau S and Ferri LE: LPS-induced TLR4 signaling in human colorectal cancer cells increases beta1 integrin-mediated cell adhesion and liver metastasis. Cancer Res. 71:1989–1998. 2011. View Article : Google Scholar : PubMed/NCBI
14	Santaolalla R, Sussman DA, Ruiz JR, Davies JM, Pastorini C, España CL, Sotolongo J, Burlingame O, Bejarano PA, Philip S, et al: TLR4 activates the β-catenin pathway to cause intestinal neoplasia. PLoS One. 8:e632982013. View Article : Google Scholar : PubMed/NCBI
15	Song W, Tiruthani K, Wang Y, Shen L, Hu M, Dorosheva O, Qiu K, Kinghorn KA, Liu R and Huang L: Trapping of lipopolysaccharide to promote immunotherapy against colorectal cancer and attenuate liver metastasis. Adv Mater. 30:e18050072018. View Article : Google Scholar : PubMed/NCBI
16	Zhao D, Sun T, Zhang X, Guo Y, Yu D, Yang M, Tan W, Wang G and Lin D: Role of CD14 promoter polymorphisms in Helicobacter pylori infection-related gastric carcinoma. Clin Cancer Res. 13:2362–2368. 2007. View Article : Google Scholar : PubMed/NCBI
17	Arihara F, Mizukoshi E, Kitahara M, Takata Y, Arai K, Yamashita T, Nakamoto Y and Kaneko S: Increase in CD14+HLA-DR-/low myeloid-derived suppressor cells in hepatocellular carcinoma patients and its impact on prognosis. Cancer Immunol Immunother. 62:1421–1430. 2013. View Article : Google Scholar : PubMed/NCBI
18	Huang A, Zhang B, Wang B, Zhang F, Fan KX and Guo YJ: Increased CD14(+)HLA-DR (−/low) myeloid-derived suppressor cells correlate with extrathoracic metastasis and poor response to chemotherapy in non-small cell lung cancer patients. Cancer Immunol Immunother. 62:1439–1451. 2013. View Article : Google Scholar : PubMed/NCBI
19	Lin Y, Dong J, Yu W, Li Y, Liu Z, Liu J, Wang C, Qin J, Zhu L and Liang J: CD14, a novel surface marker of esophageal cancer stem cells. Oncol Rep. 49:132023. View Article : Google Scholar : PubMed/NCBI
20	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. View Article : Google Scholar : PubMed/NCBI
21	Zalewski A, Snook AE and Waldman SA: Stem cells as therapeutic targets in colorectal cancer. Per Med. 18:171–183. 2021. View Article : Google Scholar : PubMed/NCBI
22	Cui G, Xu G, Zhu L, Pang Z, Zheng W, Li Z and Yuan A: Temporal and spatial changes of cells positive for stem-like markers in different compartments and stages of human colorectal adenoma-carcinoma sequence. Oncotarget. 8:45311–45322. 2017. View Article : Google Scholar : PubMed/NCBI
23	Akbarzadeh M, Maroufi NF, Tazehkand AP, Akbarzadeh M, Bastani S, Safdari R, Farzane A, Fattahi A, Nejabati HR, Nouri M and Samadi N: Current approaches in identification and isolation of cancer stem cells. J Cell Physiol. 234:14759–14772. 2019. View Article : Google Scholar : PubMed/NCBI
24	Han GD, Sun Y, Hui HX, Tao MY, Liu YQ and Zhu J: MiR-1224 acts as a prognostic biomarker and inhibits the progression of gastric cancer by targeting SATB1. Front Oncol. 11:7488962021. View Article : Google Scholar : PubMed/NCBI
25	Vargo-Gogola T and Rosen JM: Modelling breast cancer: One size does not fit all. Nat Rev Cancer. 7:659–672. 2007. View Article : Google Scholar : PubMed/NCBI
26	Tsai RY: Balancing self-renewal against genome preservation in stem cells: How do they manage to have the cake and eat it too? Cell Mol Life Sci. 73:1803–1823. 2016. View Article : Google Scholar : PubMed/NCBI
27	Chiodi I, Belgiovine C, Donà F, Scovassi AI and Mondello C: Drug treatment of cancer cell lines: A way to select for cancer stem cells? Cancers (Basel). 3:1111–1128. 2011. View Article : Google Scholar : PubMed/NCBI
28	Fang Y, Yan C, Zhao Q, Zhao B, Liao Y, Chen Y, Wang D and Tang D: The association between gut microbiota, toll-like receptors, and colorectal cancer. Clin Med Insights. 16:117955492211305492022.PubMed/NCBI
29	Liu WT, Jing YY, Yan F, Han ZP, Lai FB, Zeng JX, Yu GF, Fan QM, Li R, Zhao QD, et al: LPS-induced CXCR4-dependent migratory properties and a mesenchymal-like phenotype of colorectal cancer cells. Cell Adh Migr. 11:13–23. 2017. View Article : Google Scholar : PubMed/NCBI
30	Jeong JW, Park C, Cha HJ, Hong SH, Park SH, Kim GY, Kim WJ, Kim CH, Song KS and Choi YH: Cordycepin inhibits lipopolysaccharide-induced cell migration and invasion in human colorectal carcinoma HCT-116 cells through down-regulation of prostaglandin E2 receptor EP4. BMB Rep. 51:532–537. 2018. View Article : Google Scholar : PubMed/NCBI
31	Zhu G, Cheng Z, Lin C, Hoffman RM, Huang Y, Singh SR, Zheng W, Yang S and Ye J: MyD88 Regulates LPS-induced NF-ĸB/MAPK cytokines and promotes inflammation and malignancy in colorectal cancer cells. Cancer Genomics Proteomics. 16:409–419. 2019. View Article : Google Scholar : PubMed/NCBI
32	d'Hennezel E, Abubucker S, Murphy LO and Cullen TW: Total Lipopolysaccharide from the human gut microbiome silences toll-like receptor signaling. mSystems. 2:e00046–17. 2017.PubMed/NCBI
33	Kang M, Edmundson P, Araujo-Perez F, McCoy AN, Galanko J and Keku TO: Association of plasma endotoxin, inflammatory cytokines and risk of colorectal adenomas. BMC Cancer. 13:912013. View Article : Google Scholar : PubMed/NCBI
34	Zhu G, Huang Q, Huang Y, Zheng W, Hua J, Yang S, Zhuang J, Wang J and Ye J: Lipopolysaccharide increases the release of VEGF-C that enhances cell motility and promotes lymphangiogenesis and lymphatic metastasis through the TLR4- NF-κB/JNK pathways in colorectal cancer. Oncotarget. 7:73711–73724. 2016. View Article : Google Scholar : PubMed/NCBI
35	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. 10:87772020. View Article : Google Scholar : PubMed/NCBI
36	Zhou W, Chen X, Hu Q, Chen X, Chen Y and Huang L: Galectin-3 activates TLR4/NF-κB signaling to promote lung adenocarcinoma cell proliferation through activating lncRNA-NEAT1 expression. BMC Cancer. 18:5802018. View Article : Google Scholar : PubMed/NCBI
37	Wang W and Wang J: Toll-Like Receptor 4 (TLR4)/Cyclooxygenase-2 (COX-2) regulates prostate cancer cell proliferation, migration, and invasion by NF-κB activation. Med Sci Monit. 24:5588–5597. 2018. View Article : Google Scholar : PubMed/NCBI
38	Rakoff-Nahoum S and Medzhitov R: Regulation of spontaneous intestinal tumorigenesis through the adaptor protein MyD88. Science. 317:124–127. 2007. View Article : Google Scholar : PubMed/NCBI
39	Huang HY, Zhang ZJ, Cao CB, Wang N, Liu FF, Peng JQ, Ren XJ and Qian J: The TLR4/NF-κB signaling pathway mediates the growth of colon cancer. Eur Rev Med Pharmacol Sci. 18:3834–3843. 2014.PubMed/NCBI
40	Fukata M, Chen A, Vamadevan AS, Cohen J, Breglio K, Krishnareddy S, Hsu D, Xu R, Harpaz N, Dannenberg AJ, et al: Toll-like receptor-4 promotes the development of colitis-associated colorectal tumors. Gastroenterology. 133:1869–1881. 2007. View Article : Google Scholar : PubMed/NCBI
41	Fukata M, Chen A, Klepper A, Krishnareddy S, Vamadevan AS, Thomas LS, Xu R, Inoue H, Arditi M, Dannenberg AJ and Abreu MT: Cox-2 is regulated by Toll-like receptor-4 (TLR4) signaling: Role in proliferation and apoptosis in the intestine. Gastroenterology. 131:862–877. 2006. View Article : Google Scholar : PubMed/NCBI
42	Fukata M, Shang L, Santaolalla R, Sotolongo J, Pastorini C, España C, Ungaro R, Harpaz N, Cooper HS, Elson G, et al: Constitutive activation of epithelial TLR4 augments inflammatory responses to mucosal injury and drives colitis-associated tumorigenesis. Inflamm Bowel Dis. 17:1464–1473. 2011. View Article : Google Scholar : PubMed/NCBI
43	Cheah MT, Chen JY, Sahoo D, Contreras-Trujillo H, Volkmer AK, Scheeren FA, Volkmer JP and Weissman IL: CD14-expressing cancer cells establish the inflammatory and proliferative tumor microenvironment in bladder cancer. Proc Natl Acad Sci USA. 112:4725–4730. 2015. View Article : Google Scholar : PubMed/NCBI