CD14 regulates gastric cancer cell epithelial‑mesenchymal transition and invasion in vitro

Li,Kang; Dan,Zeng; Hu,Xuejun; Gesang,Luobu; Ze,Yongge; Bianba,Zhaxi

doi:10.3892/or.2013.2733

CD14 regulates gastric cancer cell epithelial‑mesenchymal transition and invasion in vitro

Authors:
- Kang Li
- Zeng Dan
- Xuejun Hu
- Luobu Gesang
- Yongge Ze
- Zhaxi Bianba
View Affiliations

Affiliations: Department of Gastroenterology, People's Hospital of Tibet Autonomous Region, Lhasa 850000, P.R. China, Department of Oncology, People's Hospital of Tibet Autonomous Region, Lhasa 850000, P.R. China
Published online on: September 19, 2013 https://doi.org/10.3892/or.2013.2733
Pages: 2725-2732

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

Cluster of differentiation 14 (CD14) protein functions as a co-receptor with either the Toll-like receptor TLR4 or MD-2 in the detection of bacterial lipopolysaccharide (LPS) and plays a role in the innate immune system. Recently, it was shown to have effects on the regulation of epithelial-mesenchymal transition (EMT). Thus, the present study investigated the effects of CD14 knockdown on the regulation of gastric cancer cell EMT and invasive capacity following treatment with or without LPS. Knockdown of CD14 expression using CD14 shRNA in MGC-803 cells significantly enhanced E-cadherin expression, but reduced N-cadherin and vimentin expression in both LPS-treated and untreated cells. Morphologically, the phenotype of LPS-treated CD14-knockdown cells was altered to a sporadic long spindle shape. Moreover, TNF-α-treated cells were further elongated, connections between cells were reduced, the gap between the cells was increased and the cells were transformed into mesenchymal cells. Furthermore, the invasive capacity of CD14-knockdown cells was significantly lower compared to that of the negative control shRNA-transfected MGC-803 cells. LPS-treated CD14-knockdown cells had significantly lower levels of tumor cell invasive ability when compared to the LPS-treated parental MGC-803 cells. However, addition of TNF-α to LPS-treated CD14-knockdown cells significantly increased tumor cell invasion. This study demonstrated that CD14 promoted tumor cell EMT and invasion through TNF-α, whereas knockdown of CD14 expression inhibited gastric cancer cell invasion and EMT.

View Figures

View References

1	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
2	Ji J: Chinese guidelines for diagnosis and treatment of gastric cancer (2011 edition). Transl Gastrointest Cancer. 1:103–114. 2012.
3	Thiery JP: Epithelial-mesenchymal transitions in tumor progression. Nat Rev Cancer. 2:442–454. 2002. View Article : Google Scholar : PubMed/NCBI
4	Herrera V and Parsonnet J: Helicobacter pylori and gastric adenocarcinoma. Clin Microbiol Infect. 15:971–976. 2009. View Article : Google Scholar
5	Wright SD, Ramos RA, Tobias PS, Ulevitch RJ and Mathison JC: CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science. 249:1431–1433. 1990. View Article : Google Scholar : PubMed/NCBI
6	Pugin J, Heumann ID, Tomasz A, Kravchenko VV, Akamatsu Y, Nishijima M, Glauser MP, Tobias PS and Ulevitch RJ: CD14 is a pattern recognition receptor. Immunity. 1:509–516. 1994. View Article : Google Scholar : PubMed/NCBI
7	Chen MC, Chang WW, Kuan YD, Lin ST, Hsu HC and Lee CH: Resveratrol inhibits LPS-induced epithelial-mesenchymal transition in mouse melanoma model. Innate Immun. 18:685–693. 2012. View Article : Google Scholar : PubMed/NCBI
8	Gioannini TL and Weiss JP: Regulation of interactions of Gram-negative bacterial endotoxins with mammalian cells. Immunol Res. 39:249–260. 2007. View Article : Google Scholar : PubMed/NCBI
9	Miyake K: Innate immune sensing of pathogens and danger signals by cell surface Toll-like receptors. Semin Immunol. 19:3–10. 2007. View Article : Google Scholar : PubMed/NCBI
10	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
11	Kanczkowski W, Tymoszuk P, Ehrhart-Bornstein M, Wirth MP, Zacharowski K and Bornstein SR: Abrogation of TLR4 and CD14 expression and signaling in human adrenocortical tumors. J Clin Endocrinol Metab. 95:E421–E429. 2010. View Article : Google Scholar : PubMed/NCBI
12	Zanoni I, Ostuni R, Capuano G, Collini M, Caccia M, Ronchi AE, Rocchetti M, Mingozzi F, Foti M, Chirico G, Costa B, Zaza A, Ricciardi-Castagnoli P and Granucci F: CD14 regulates the dendritic cell life cycle after LPS exposure through NFAT activation. Nature. 460:264–268. 2009. View Article : Google Scholar : PubMed/NCBI
13	Reynolds A, Leake D, Boese Q, Scaringe S, Marshall WS and Khvorova A: Rational siRNA design for RNA interference. Nat Biotechnol. 22:326–330. 2004. View Article : Google Scholar : PubMed/NCBI
14	Kawata M, Koinuma D, Ogami T, Umezawa K, Iwata C, Watabe T and Miyazono K: TGF-β-induced epithelial-mesenchymal transition of A549 lung adenocarcinoma cells is enhanced by pro-inflammatory cytokines derived from RAW 264.7 macrophage cells. J Biochem. 151:205–216. 2012.
15	Bates RC and Mercurio AM: Tumor necrosis factor-α stimulates the epithelial-to-mesenchymal transition of human colonic organoids. Mol Biol Cell. 14:1790–1800. 2003.
16	Guarino M: Epithelial-mesenchymal transition and tumour invasion. Int J Biochem Cell Biol. 39:2153–2160. 2007. View Article : Google Scholar : PubMed/NCBI
17	Nakajima S, Doi R, Toyoda E, Tsuji S, Wada M, Koizumi M, Tulachan SS, Ito D, Kami K, Mori T, Kawaguchi Y, Fujimoto K, Hosotani R and Imamura M: N-cadherin expression and epithelial-mesenchymal transition in pancreatic carcinoma. Clin Cancer Res. 10:4125–4133. 2004. View Article : Google Scholar : PubMed/NCBI
18	Cavallaro U, Schaffhauser B and Christofori G: Cadherins and the tumour progression: is it all in a switch? Cancer Lett. 176:123–128. 2002. View Article : Google Scholar : PubMed/NCBI
19	Lang SH, Hyde C, Reid IN, Hitchcock IS, Hart CA, Bryden AA, Villette JM, Stower MJ and Maitland NJ: Enhanced expression of vimentin in motile prostate cell lines and in poorly differentiated and metastatic prostate carcinoma. Prostate. 52:253–263. 2002. View Article : Google Scholar : PubMed/NCBI
20	Polk DB and Peek RM Jr: Helicobacter pylori: gastric cancer and beyond. Nature Rev Cancer. 10:403–414. 2010. View Article : Google Scholar
21	Triantafilou M and Triantafilou K: The dynamics of LPS recognition: complex orchestration of multiple receptors. J Endotoxin Res. 11:5–11. 2005.PubMed/NCBI
22	Le Roy D, Di Padova F, Adachi Y, Glauser MP, Calandra T and Heumann D: Critical role of lipopolysaccharide-binding protein and CD14 in immune responses against gram-negative bacteria. J Immunol. 167:2759–2765. 2001.PubMed/NCBI
23	Schimke J, Mathison J, Morgiewicz J and Ulevitch RJ: Anti-CD14 mAb treatment provides therapeutic benefit after in vivo exposure to endotoxin. Proc Natl Acad Sci USA. 95:13875–13880. 1998. View Article : Google Scholar : PubMed/NCBI
24	Wu Y, Deng J, Rychahou PG, Qiu S, Evers BM and Zhou BP: Stabilization of snail by NF-κB is required for inflammation-induced cell migration and invasion. Cancer Cell. 15:416–428. 2009.
25	Iwano M: EMT and TGF-beta in renal fibrosis. Front Biosci. 2:229–238. 2010. View Article : Google Scholar : PubMed/NCBI
26	Zavadil J and Böttinger EP: TGF-β and epithelial-to-mesenchymal transitions. Oncogene. 24:5764–5774. 2005.
27	Seki E, De Minicis S, Osterreicher CH, Kluwe J, Osawa Y, Brenner DA and Schwabe RF: TLR4 enhances TGF-β signaling and hepatic fibrosis. Nat Med. 13:1324–1332. 2007.
28	He W, Liu Q, Wang L, Chen W, Li N and Cao X: TLR4 signaling promotes immune escape of human lung cancer cells by inducing immunosuppressive cytokines and apoptosis resistance. Mol Immunol. 44:2850–2859. 2007. View Article : Google Scholar : PubMed/NCBI
29	Song MN and Cho SY: CD14 acts as an angiogenic factor by inducing basic fibroblast growth factor (bFGF). Bull Korean Chem Soc. 28:1613–1614. 2007. View Article : Google Scholar
30	Hua D, Liu MY, Cheng ZD, Qin XJ, Zhang HM, Chen Y, Qin GJ, Liang G, Li JN, Han XF and Liu DX: Small interfering RNA-directed targeting of Toll-like receptor 4 inhibits human prostate cancer cell invasion, survival and tumorigenicity. Mol Immunol. 46:2876–2884. 2009. View Article : Google Scholar : PubMed/NCBI