The Expression And Significance Of Toll-like Receptor2,3,4on Head And Neck Squamous Cells(HNSCs)

ObjectsLaryngocarcinoma or hypopharyngeal carcinoma, the most common pathological type ofwhich is the squamous cell carcinoma, shows high degree of malignancy account for1-5%of themalignant tumor and is often with lymph node metastases around the neck. Surgery operation isthe main therapeutic method for this severe disease. Throat is often in a inflammation situationbecause of direct contact with environment which leads to physical or chemical stimulation.Chronic inflammation plays a significant role in the process of tumorgenesis.Toll-like receptor is an important pattern recognition receptor that recognizes highconservative molecular structure, pathogen-associated molecular patterns and danger-associatedmolecular patterns. TLRs is a kind of classic transmembrane protein. As one of the mostcharacteristic PPR molecule, it can recruit several adaptor protein and trigger signal transduction,activate the specific transcription factors, lead to antibacterial peptide, pro-inflammatorycytokines and chemokines,antiviral agent, adhesion molecule and other active factors.Research results demonstrate that TLRs expresses highly not only in immune cells, but alsoin many kinds of malignancy cells, especially in inflammatory related tumors. It can exert animportant effect in the process of tumor and treatment. This research will study the expressionintensity of TLR2,3and4at transcription and protein levels, using their correspondingpara-carcinoma tissue and polyp tissue as control. On the basis of our results, to discuss the influence of TLR2,3and4in cancer process of Laryngocarcinoma or hypopharyngealcarcinoma refering to other findings. We hope to offer more ideas about cancer dignosis,prevention and treatment.Methods and results1．Detecting the expression levels of TLR2,3,4mRNA in diffrent type pathological tissue byRT-PCR.1.1The diffrent expression level of TLR2,3,4mRNA in tumor tissue and vocal polyps.TLR2,3,4mRNA expressed in tumor tissue, but fairly expressed in the vocal polyps. TLR4isthe top one, followed by TLR2. TLRs expression level in tumor tissue was stronger than that inthe vocal polyps. TLR2t=6.723，TLR3t=4.976, TLR4t=7.645，all P<0.01. This statistic resulthas great significance.1.2The diffrent expression levels of TLR2,3,4mRNA in tumor tissue and its correspondingadjacent tissue of cancer.TLR2,3,4mRNA expressed both in tumor and the adjacent tissue of cancer. Their expressionlevels in tumor tissue was stronger than that in the adjacent tissue of tumor. TLR2t=6.179，TLR3t=4.831, TLR4t=6.057，all P<0.01. This statistic result has great significance.1.3The diffrent expression levels of TLR2,3,4mRNA in different tumor stages.TLR2,3,4mRNA have great significance in different tumor clinical and TNM stages and theirexpression levels were closely associated with the tumor stages. ANOVA results: in differenttumor clinical stages: TLR2F=5.150，TLR3F=3.700，TLR4F=4.862，all P <0.05. TNM(N)stage：TLR2F=5.562，TLR3F=7.200，TLR4F=15.039，all P <0.05. All of these statisticresults have great significance.1.4The diffrent expression levels of TLR2,3,4mRNA in different pathological grade of cancer.The expression levels of TLR2,4mRNA is positive associated with its grade malignancy,while the expression intensity of TLR3mRNA may has not significance with it in cancer tissue. 2．Detecting the expression levels of TLR2,4protein in diffrent types of pathological tissue byWesteron blot.2.1The diffrent expression levels of TLR2,4protein in cancer tissue and the vocal polyps.Protein expression levels of TLR2,4in tumor tissue were stronger than that in the vocalpolyps. Especially that TLR2showed the more difference between in these two kinds of tissue.2.2The diffrent expression levels of TLR2、4proteins in cancer tissue and the adjacent tissueof cancer.Protein expression levels of TLR2,4in cancer tissue were stronger than that in the adjacenttissue of cancer.2.3The diffrent expression levels of TLR2,4protein in different tumor stages.TLR2、4protein have great significance in different tumor clinical and TNM stages and theirexpression levels were closely associated with the tumor stages.2.4The diffrent expression levels of TLR2,4protein in different pathological grade of cancer.Threre were positive correlation between protein expression levels of TLR2,4and themalignant degree of tumor.3. Detecting the expression locations of TLR2,4protein in the squamous carcinoma cells byimmunohistochemistry.Both TLR2,4expressed on cell membrane and they also can be seen in cytoplasm of thesquamous carcinoma cells. Anti-TLR2,4Ab，positive cells showing cytoplasmic and membranebound reaction.Conclusion: TLR2,3,4expressed usually in the Laryngocarcinoma or hypopharyngealcarcinoma tissue but fairly seen in the vocal polyps. Both TLR2,4expressed on cell membraneand they also can be seen in cytoplasm of the squamous carcinoma cells. Expression levels ofTLR2,3,4mRNA or protein showed a great significance between in the cancer tissue and theircorresponding adjacent tissue of cancer. The expression levels of TLR2,4mRNA is positiveassociated with its grade malignancy, while the expression intensity of TLR3mRNA might has not significance with it in cancer tissue.Significance of researchThis project confirmed the expression levels of TLR2、3and4mRNA and protein levels ofTLR2and4as well as cellular localization in Laryngocarcinoma or hypopharyngeal carcinomatissue. We also make sure the relation about expression intensity of these TLRs with tumor stagesand lymph node metastasis. This can offer theory evidence for the cilinic application.