Effect of TLR4 on the growth of SiHa human cervical cancer cells via the MyD88‑TRAF6-TAK1 and NF-κB-cyclin D1-STAT3 signaling pathways

Ma,Li; Feng,Li; Ding,Xiaoping; Li,Yongwang

doi:10.3892/ol.2018.7801

Effect of TLR4 on the growth of SiHa human cervical cancer cells via the MyD88‑TRAF6-TAK1 and NF-κB-cyclin D1-STAT3 signaling pathways

Authors:
- Li Ma
- Li Feng
- Xiaoping Ding
- Yongwang Li
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, The Second Artillery General Hospital, Beijing 100088, P.R. China, Department of Anesthesiology, The Second Artillery General Hospital, Beijing 100088, P.R. China
Published online on: January 16, 2018 https://doi.org/10.3892/ol.2018.7801
Pages: 3965-3970

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

The present study aimed to investigate the effect of Toll‑like receptor 4 (TLR4) on SiHa human cervical cancer cells and its potential molecular biological mechanisms. The expression of TLR4 following treatment with lipopolysaccharide (LPS) in in SiHa cervical cancer cells was detected by quantitative polymerase chain reaction (qPCR). LPS‑induced cell proliferation and apoptosis were detected by MTT assay as well as staining with propidium iodide (PI) and Annexin V/PI double staining. qPCR was performed to analyze the expression levels of tumor necrosis factor receptor‑associated factor 6 (TRAF6) and transforming growth factor‑activated kinase 1 (TAK1) genes. Western blot analysis was performed to analyze the expression of myeloid differentiation 88 (MyD88), nuclear factor‑κB (NF‑κB), cyclin D1 and signal transducer and activator of transcription 3 (STAT3) proteins. In the present study, it was revealed that TLR4 expression in SiHa cervical cancer cells may be upregulated by LPS. Additionally, LPS was able to increase the proliferation of SiHa cells. However, LPS treatment did not have an effect on apoptosis of the cells. In addition, the MyD88‑TRAF6‑TAK1 and NF‑κB‑cyclin D1‑STAT3 signaling pathways were induced in SiHa cells by LPS. These results suggested the effect of LPS and TLR4 on proliferation of SiHa human cervical cancer cells via the MyD88‑TRAF6‑TAK1 and NF‑κB‑cyclin D1-STAT3 signaling pathways.

View Figures

View References

1	Symonds RP, Gourley C, Davidson S, Carty K, McCartney E, Rai D, Banerjee S, Jackson D, Lord R, McCormack M, et al: Cediranib combined with carboplatin and paclitaxel in patients with metastatic or recurrent cervical cancer (CIRCCa): A randomised, double-blind, placebo-controlled phase 2 trial. Lancet Oncol. 16:1515–1524. 2015. View Article : Google Scholar : PubMed/NCBI
2	Braicu EI, Fotopoulou C, Chekerov R, Richter R, Blohmer J, Kümmel S, Stamatian F, Yalcinkaya I, Mentze M, Lichtenegger W and Sehouli J: Role of serum concentration of VEGFR1 and TIMP2 on clinical outcome in primary cervical cancer: Results of a companion protocol of the randomized, NOGGO-AGO phase III adjuvant trial of simultaneous cisplatin-based radiochemotherapy vs. carboplatin and paclitaxel containing sequential radiotherapy. Cytokine. 61:755–758. 2013. View Article : Google Scholar : PubMed/NCBI
3	Burtness B, Bourhis JP, Vermorken JB, Harrington KJ and Cohen EE: Afatinib versus placebo as adjuvant therapy after chemoradiation in a double-blind, phase III study (LUX-Head & Neck 2) in patients with primary unresected, clinically intermediate-to-high-risk head and neck cancer: Study protocol for a randomized controlled trial. Trials. 15:4692014. View Article : Google Scholar : PubMed/NCBI
4	Muwonge R, Wesley RS, Nene BM, Shastri SS, Jayant K, Malvi SG, Thara S and Sankaranarayanan R: Evaluation of cytology and visual triage of human papillomavirus-positive women in cervical cancer prevention in India. Int J Cancer. 134:2902–2909. 2014. View Article : Google Scholar : PubMed/NCBI
5	DeCarlo CA, Rosa B, Jackson R, Niccoli S, Escott NG and Zehbe I: Toll-like receptor transcriptome in the HPV-positive cervical cancer microenvironment. Clin Dev Immunol. 2012:7858252012. View Article : Google Scholar : PubMed/NCBI
6	Werner J, Decarlo CA, Escott N, Zehbe I and Ulanova M: Expression of integrins and Toll-like receptors in cervical cancer: Effect of infectious agents. Innate Immun. 18:55–69. 2012. View Article : Google Scholar : PubMed/NCBI
7	Li L, Cheng FW, Wang F, Jia B, Luo X and Zhang SQ: The activation of TLR7 regulates the expression of VEGF, TIMP1, MMP2, IL-6, and IL-15 in Hela cells. Mol Cell Biochem. 389:43–49. 2014. View Article : Google Scholar : PubMed/NCBI
8	Zhan Z, Xie X, Cao H, Zhou X, Zhang XD, Fan H and Liu Z: Autophagy facilitates TLR4- and TLR3-triggered migration and invasion of lung cancer cells through the promotion of TRAF6 ubiquitination. Autophagy. 10:257–268. 2014. View Article : Google Scholar : PubMed/NCBI
9	Xiao J, Guo Q, Wang X, Xie F, Zhang H and Sui L: Study on the expression and signification of TLR4/NO pathway in cervical tumorigenesis with high risk HPV infection. Zhonghua Fu Chan Ke Za Zhi. 50:41–47. 2015.(In Chinese). PubMed/NCBI
10	He A, Ji R, Shao J, He C, Jin M and Xu Y: TLR4-MyD88-TRAF6-TAK1 complex-mediated NF-κB activation contribute to the anti-inflammatory effect of V8 in LPS-induced human cervical cancer SiHa cells. Inflammation. 39:172–181. 2016. View Article : Google Scholar : PubMed/NCBI
11	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
12	Kirchheiner K, Nout RA, Czajka-Pepl A, Ponocny-Seliger E, Sturdza AE, Dimopoulos JC, Dörr W and Pötter R: Health related quality of life and patient reported symptoms before and during definitive radio (chemo)therapy using image-guided adaptive brachytherapy for locally advanced cervical cancer and early recovery-a mono-institutional prospective study. Gynecol Oncol. 136:415–423. 2015. View Article : Google Scholar : PubMed/NCBI
13	Tomomura M, Suzuki R, Shirataki Y, Sakagami H, Tamura N and Tomomura A: Rhinacanthin C inhibits osteoclast differentiation and bone resorption: Roles of TRAF6/TAK1/MAPKs/NF-κB/NFATc1 signaling. PLoS One. 10:e01301742015. View Article : Google Scholar : PubMed/NCBI
14	He W and Cronstein BN: Adenosine A1 receptor regulates osteoclast formation by altering TRAF6/TAK1 signaling. Purinergic Signal. 8:327–337. 2012. View Article : Google Scholar : PubMed/NCBI
15	Bai C, Yang X, Zou K, He H, Wang J, Qin H, Yu X, Liu C, Zheng J, Cheng F and Chen J: Anti-proliferative effect of RCE-4 from Reineckia carnea on human cervical cancer HeLa cells by inhibiting the PI3K/Akt/mTOR signaling pathway and NF-κB activation. Naunyn Schmiedebergs Arch Pharmacol. 389:573–584. 2016. View Article : Google Scholar : PubMed/NCBI
16	Zeng KW, Yu Q, Liao LX, Song FJ, Lv HN, Jiang Y and Tu PF: Anti-neuroinflammatory effect of MC13, a novel coumarin compound from condiment murraya, through inhibiting lipopolysaccharide-induced TRAF6-TAK1-NF-κB, P38/ERK MAPKS and Jak2-Stat1/Stat3 pathways. J Cell Biochem. 116:1286–1299. 2015. View Article : Google Scholar : PubMed/NCBI
17	Nees M, Geoghegan JM, Hyman T, Frank S, Miller L and Woodworth CD: Papillomavirus type 16 oncogenes downregulate expression of interferon-responsive genes and upregulate proliferation-associated and NF-kappaB-responsive genes in cervical keratinocytes. J Virol. 75:4283–4296. 2001. View Article : Google Scholar : PubMed/NCBI
18	Yang L, Gu L, Li Z and Zhou M: Translation of TRAF1 is regulated by IRES-dependent mechanism and stimulated by vincristine. Nucleic Acids Res. 38:4503–4513. 2010. View Article : Google Scholar : PubMed/NCBI
19	Capalbo G, Mueller-Kuller T, Koschmieder S, Klein HU, Ottmann OG, Hoelzer D and Scheuring UJ: Characterization of ZC3H15 as a potential TRAF-2-interacting protein implicated in the NFκB pathway and overexpressed in AML. Int J Oncol. 43:246–254. 2013. View Article : Google Scholar : PubMed/NCBI
20	Wu Y, Fu H, Zhang H, Huang H, Chen M, Zhang L, Yang H and Qin D: Cyclin D1 (CCND1) G870A polymorphisms and cervical cancer susceptibility: A meta-analysis based on ten case-control studies. Tumour Biol. 35:6913–6918. 2014. View Article : Google Scholar : PubMed/NCBI
21	Ni HJ, Chang YN, Kao PH, Chai SP, Hsieh YH, Wang DH and Fong JC: Depletion of SUMO ligase hMMS21 impairs G1 to S transition in MCF-7 breast cancer cells. Biochim Biophys Acta. 1820:1893–1900. 2012. View Article : Google Scholar : PubMed/NCBI
22	Chen J, Bai M, Ning C, Xie B, Zhang J, Liao H, Xiong J, Tao X, Yan D, Xi X, et al: Gankyrin facilitates follicle-stimulating hormone-driven ovarian cancer cell proliferation through the PI3K/AKT/HIF-1α/cyclin D1 pathway. Oncogene. 35:2506–2517. 2016. View Article : Google Scholar : PubMed/NCBI
23	Fan Z, Cui H, Yu H, Ji Q, Kang L, Han B, Wang J, Dong Q, Li Y, Yan Z, et al: MiR-125a promotes paclitaxel sensitivity in cervical cancer through altering STAT3 expression. Oncogenesis. 5:e1972016. View Article : Google Scholar : PubMed/NCBI
24	Zhang P, Yang B, Yao YY, Zhong LX, Chen XY, Kong QY, Wu ML, Li C, Li H and Liu J: PIAS3, SHP2 and SOCS3 expression patterns in cervical cancers: Relevance with activation and resveratrol-caused inactivation of STAT3 signaling. Gynecol Oncol. 139:529–535. 2015. View Article : Google Scholar : PubMed/NCBI