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TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention

  • Authors:
    • Xiao Yang
    • Gan Tao Chen
    • Yan Qing Wang
    • Shu Xian
    • Li Zhang
    • Shao Ming Zhu
    • Feng Pan
    • Yan Xiang Cheng
  • View Affiliations / Copyright

    Affiliations: Department of Obstetrics and Gynecology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Gastroenterology, The Third Renmin Hospital of Xiantao City, Xiantao, Hubei 433000, P.R. China, Department of Urology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China
    Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 2229-2238
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    Published online on: November 20, 2017
       https://doi.org/10.3892/mmr.2017.8108
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Abstract

The present study investigated the mechanism underlying Toll-like receptor 4 (TLR4)-mediated stimulation of hypoxia-inducible factor-1α (HIF-1α) activity and its association with reactive oxygen species (ROS) in cervical cancer cells. SiHa cells were cultured and randomized to control, lipopolysaccharide (LPS), methyl-β-cyclodextrin (MβCD)+LPS, ammonium pyrrolidinedithiocarbamate (PDTC)+LPS, ST2825+LPS and small interfering (si) RNA TLR4+LPS treatment groups. Cell proliferation was quantified using an MTT assay, cell cloning was performed using soft agar colony formation and HIF-1α expression was detected by immunocytochemical staining and western blot analyses. Dichloro-dihydro-fluorescein diacetate and lucigenin luminescence assays were used to detect alterations in ROS and nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase content, respectively. Co-localization of TLR4 and HIF-1α was detected by immunofluorescence staining and observed using fluorescence microscopy. Compared with the control group, cell proliferation was enhanced in the LPS-treated group and was not altered in the PDTC+LPS treatment group. Cell proliferation was reduced in all other treatment groups (P<0.05). Compared with the LPS group, cell proliferation decreased in all other groups. Compared with the PDTC+LPS treatment group, cell proliferation significantly decreased when LPS was co-administered with ST2825, siTLR4 and MβCD (P<0.01). Treatment with MβCD+LPS exhibited an increased inhibitory effect on cell activity and proliferation. Compared with the control group, HIF-1α expression was enhanced following treatment with LPS, although it decreased when LPS was co-administered with ST2825, siTLR4 and MβCD (P<0.05). HIF-1α expression decreased following treatment with ST2825, siTLR4, MβCD and PDTC+LPS, compared with treatment with LPS alone. Compared with the PDTC+LPS group, HIF-1α activity decreased when LPS was co-administered with ST2825, siTLR4 and MβCD. NADPH oxidase and ROS levels increased in cells treated with LPS, compared with the control group, at 24 and 12 h following treatment, respectively, and decreased at 12 h when LPS was co-administered with ST2825, siTLR4 and MβCD. There was no difference between the LPS and PDTC+LPS groups with respect to NADPH and ROS levels. Compared with the PDTC+LPS group, NADPH oxidase activity and ROS content decreased when LPS was co-administered with ST2825, siTLR4 and MβCD. NADPH oxidase activity and ROS content were lowest in the MβCD+LPS treatment group, and immunofluorescent staining demonstrated that TLR4 was localized to the cell surface and HIF-1α was primarily localized to the cytoplasm. TLR4 was co-expressed with HIF-1α in cervical cancer cells. The results of the present study suggested that TLR4 signaling primarily promoted HIF-1α activity via activation of lipid rafts/NADPH oxidase redox signaling and may be associated with the initiation and progression of cervical cancer. This promoting effect was stronger in TLR4/lipid rafts/NADPH oxidase pathway than that in TLR4-NF-κB signaling pathway. Therefore, the TLR4/lipid raft-associated redox signal may be a target for therapeutic intervention to prevent the growth of cervical cancer.
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Copy and paste a formatted citation
Spandidos Publications style
Yang X, Chen GT, Wang YQ, Xian S, Zhang L, Zhu SM, Pan F and Cheng YX: TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention. Mol Med Rep 17: 2229-2238, 2018.
APA
Yang, X., Chen, G.T., Wang, Y.Q., Xian, S., Zhang, L., Zhu, S.M. ... Cheng, Y.X. (2018). TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention. Molecular Medicine Reports, 17, 2229-2238. https://doi.org/10.3892/mmr.2017.8108
MLA
Yang, X., Chen, G. T., Wang, Y. Q., Xian, S., Zhang, L., Zhu, S. M., Pan, F., Cheng, Y. X."TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention". Molecular Medicine Reports 17.2 (2018): 2229-2238.
Chicago
Yang, X., Chen, G. T., Wang, Y. Q., Xian, S., Zhang, L., Zhu, S. M., Pan, F., Cheng, Y. X."TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention". Molecular Medicine Reports 17, no. 2 (2018): 2229-2238. https://doi.org/10.3892/mmr.2017.8108
Copy and paste a formatted citation
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Spandidos Publications style
Yang X, Chen GT, Wang YQ, Xian S, Zhang L, Zhu SM, Pan F and Cheng YX: TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention. Mol Med Rep 17: 2229-2238, 2018.
APA
Yang, X., Chen, G.T., Wang, Y.Q., Xian, S., Zhang, L., Zhu, S.M. ... Cheng, Y.X. (2018). TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention. Molecular Medicine Reports, 17, 2229-2238. https://doi.org/10.3892/mmr.2017.8108
MLA
Yang, X., Chen, G. T., Wang, Y. Q., Xian, S., Zhang, L., Zhu, S. M., Pan, F., Cheng, Y. X."TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention". Molecular Medicine Reports 17.2 (2018): 2229-2238.
Chicago
Yang, X., Chen, G. T., Wang, Y. Q., Xian, S., Zhang, L., Zhu, S. M., Pan, F., Cheng, Y. X."TLR4 promotes the expression of HIF-1α by triggering reactive oxygen species in cervical cancer cells in vitro-implications for therapeutic intervention". Molecular Medicine Reports 17, no. 2 (2018): 2229-2238. https://doi.org/10.3892/mmr.2017.8108
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