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The influence of TLR4 agonist lipopolysaccharides on hepatocellular carcinoma cells and the feasibility of its application in treating liver cancer.

Gu J, Sun R, Shen S, Yu Z - Onco Targets Ther (2015)

Bottom Line: Fluorescent quantitative polymerase chain reaction and Western blot method were used to determine the change of expression of Cyclin D1.The results demonstrated that most TLRs were expressed in liver cancer cells; stimulating TLR4 by LPS could upregulate TLR4 mRNA and the protein level, activate NF-κB signaling pathway downstream of TLR4, and mediate the generation of inflammatory factors IL-6, IL-8, and TNF-α; LPS was found to be able to strengthen the proliferation ability of liver cancer cells, especially H7402 cells; the expression of Cyclin D1 rose and H7402 cells were promoted to transit from G1 stage to S stage under the stimulation of LPS, but cell apoptosis was not affected.It was also found that LPS was able to activate signal transducer and activator of transcription -3 (STAT3) signaling pathway in H7402 cells and meanwhile significantly increase the initiation activity of STAT3; proliferation promoting effect of LPS to liver cancer cells remarkably lowered once STAT3 was blocked or inhibited.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases, The First Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan, People's Republic of China.

ABSTRACT

Objective: This study was designed to explore the influence of Toll-like receptor 4 (TLR4) agonist lipopolysaccharides (LPS) on liver cancer cell and the feasibility to perform liver cancer adjuvant therapy.

Methods: Human liver cancer cell lines HepG2, H7402, and PLC/PRF/5 were taken as models, and the expression of TLRs mRNA was detected by real time-polymerase chain reaction method semiquantitatively. WST-1 method was used to detect the influence of LPS on the proliferation ability of liver cancer cells; propidium iodide (PI) single staining and Annexin V/PI double staining were used to test the influence of LPS on the cell cycle and apoptosis, respectively, on human liver cancer cell line H7402. Fluorescent quantitative polymerase chain reaction and Western blot method were used to determine the change of expression of Cyclin D1.

Results: The results demonstrated that most TLRs were expressed in liver cancer cells; stimulating TLR4 by LPS could upregulate TLR4 mRNA and the protein level, activate NF-κB signaling pathway downstream of TLR4, and mediate the generation of inflammatory factors IL-6, IL-8, and TNF-α; LPS was found to be able to strengthen the proliferation ability of liver cancer cells, especially H7402 cells; the expression of Cyclin D1 rose and H7402 cells were promoted to transit from G1 stage to S stage under the stimulation of LPS, but cell apoptosis was not affected. It was also found that LPS was able to activate signal transducer and activator of transcription -3 (STAT3) signaling pathway in H7402 cells and meanwhile significantly increase the initiation activity of STAT3; proliferation promoting effect of LPS to liver cancer cells remarkably lowered once STAT3 was blocked or inhibited.

Conclusion: Thus, TLR4 agonist LPS is proved to be able to induce liver cancer cells to express inflammation factors and mediate liver cancer cell proliferation and generation of multidrug resistance by activating the cyclooxygenase-2/prostaglandin signal axis as well as the STAT3 pathway.

No MeSH data available.


Related in: MedlinePlus

NF-κB signaling was activated in response to LPS stimulation.Notes: (A and B) H7402 and HepG2 cells were treated with LPS (10 μg/mL) for different time. Then total proteins were extracted. Western blot was used to detect the activation of NF-κB signal pathway.Abbreviations: LPS, lipopolysaccharides; min, minutes.
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f2-ott-8-2215: NF-κB signaling was activated in response to LPS stimulation.Notes: (A and B) H7402 and HepG2 cells were treated with LPS (10 μg/mL) for different time. Then total proteins were extracted. Western blot was used to detect the activation of NF-κB signal pathway.Abbreviations: LPS, lipopolysaccharides; min, minutes.

Mentions: Under the stimulation of inherent components related to pathogens, TLR4 started signal transduction to mediate the activation of Myd88-dependent NF-κB and mitogen-activated protein kinase (MAPK) pathway as well as Myd88-independent IRF pathway, in which NF-κB pathway is particularly important. The activation of NF-κB was detected after H7402 and HepG2 cells were stimulated with LPS (10 μg/mL) at different time points. Figure 2 shows that LPS was able to activate NF-κB signal pathway in HCC.


The influence of TLR4 agonist lipopolysaccharides on hepatocellular carcinoma cells and the feasibility of its application in treating liver cancer.

Gu J, Sun R, Shen S, Yu Z - Onco Targets Ther (2015)

NF-κB signaling was activated in response to LPS stimulation.Notes: (A and B) H7402 and HepG2 cells were treated with LPS (10 μg/mL) for different time. Then total proteins were extracted. Western blot was used to detect the activation of NF-κB signal pathway.Abbreviations: LPS, lipopolysaccharides; min, minutes.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4551308&req=5

f2-ott-8-2215: NF-κB signaling was activated in response to LPS stimulation.Notes: (A and B) H7402 and HepG2 cells were treated with LPS (10 μg/mL) for different time. Then total proteins were extracted. Western blot was used to detect the activation of NF-κB signal pathway.Abbreviations: LPS, lipopolysaccharides; min, minutes.
Mentions: Under the stimulation of inherent components related to pathogens, TLR4 started signal transduction to mediate the activation of Myd88-dependent NF-κB and mitogen-activated protein kinase (MAPK) pathway as well as Myd88-independent IRF pathway, in which NF-κB pathway is particularly important. The activation of NF-κB was detected after H7402 and HepG2 cells were stimulated with LPS (10 μg/mL) at different time points. Figure 2 shows that LPS was able to activate NF-κB signal pathway in HCC.

Bottom Line: Fluorescent quantitative polymerase chain reaction and Western blot method were used to determine the change of expression of Cyclin D1.The results demonstrated that most TLRs were expressed in liver cancer cells; stimulating TLR4 by LPS could upregulate TLR4 mRNA and the protein level, activate NF-κB signaling pathway downstream of TLR4, and mediate the generation of inflammatory factors IL-6, IL-8, and TNF-α; LPS was found to be able to strengthen the proliferation ability of liver cancer cells, especially H7402 cells; the expression of Cyclin D1 rose and H7402 cells were promoted to transit from G1 stage to S stage under the stimulation of LPS, but cell apoptosis was not affected.It was also found that LPS was able to activate signal transducer and activator of transcription -3 (STAT3) signaling pathway in H7402 cells and meanwhile significantly increase the initiation activity of STAT3; proliferation promoting effect of LPS to liver cancer cells remarkably lowered once STAT3 was blocked or inhibited.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases, The First Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan, People's Republic of China.

ABSTRACT

Objective: This study was designed to explore the influence of Toll-like receptor 4 (TLR4) agonist lipopolysaccharides (LPS) on liver cancer cell and the feasibility to perform liver cancer adjuvant therapy.

Methods: Human liver cancer cell lines HepG2, H7402, and PLC/PRF/5 were taken as models, and the expression of TLRs mRNA was detected by real time-polymerase chain reaction method semiquantitatively. WST-1 method was used to detect the influence of LPS on the proliferation ability of liver cancer cells; propidium iodide (PI) single staining and Annexin V/PI double staining were used to test the influence of LPS on the cell cycle and apoptosis, respectively, on human liver cancer cell line H7402. Fluorescent quantitative polymerase chain reaction and Western blot method were used to determine the change of expression of Cyclin D1.

Results: The results demonstrated that most TLRs were expressed in liver cancer cells; stimulating TLR4 by LPS could upregulate TLR4 mRNA and the protein level, activate NF-κB signaling pathway downstream of TLR4, and mediate the generation of inflammatory factors IL-6, IL-8, and TNF-α; LPS was found to be able to strengthen the proliferation ability of liver cancer cells, especially H7402 cells; the expression of Cyclin D1 rose and H7402 cells were promoted to transit from G1 stage to S stage under the stimulation of LPS, but cell apoptosis was not affected. It was also found that LPS was able to activate signal transducer and activator of transcription -3 (STAT3) signaling pathway in H7402 cells and meanwhile significantly increase the initiation activity of STAT3; proliferation promoting effect of LPS to liver cancer cells remarkably lowered once STAT3 was blocked or inhibited.

Conclusion: Thus, TLR4 agonist LPS is proved to be able to induce liver cancer cells to express inflammation factors and mediate liver cancer cell proliferation and generation of multidrug resistance by activating the cyclooxygenase-2/prostaglandin signal axis as well as the STAT3 pathway.

No MeSH data available.


Related in: MedlinePlus