Application of TLR3 inhibitor to preparation of product for preventing and treating neoplasm metastasis
A technology of tumor metastasis and inhibition, applied in the fields of biotechnology and medicine
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Embodiment 1
[0088] Example 1: The expression of TLR3 in the paracancerous tissue of lung cancer is related to the infiltration of neutrophils
[0089] The samples used were tissue sections of 90 lung cancer patients in 2004 (obtained from Shanghai Xinchao Biotechnology Co., Ltd.). The basic information of the patients is shown in Table 1.
[0090] Table 1. Clinical information of 90 lung cancer patients
[0091]
[0092] Immunohistochemical method: take mouse lung tissue and fix overnight with 4% paraformaldehyde. Sectioned after paraffin embedding. Bake slices at 70°C for 3 hours. After dewaxing and placing in water, antigen retrieval was carried out with citric acid. Serum was blocked for 40 minutes at 37°C. Wash 3 times with PBS. Primary antibody incubation was incubated overnight at 4°C. Wash 3 times with PBS. Secondary antibody incubation was incubated at 37°C for 30 minutes. Wash 3 times with PBS. Horseradish peroxidase was incubated at 37°C for 20 minutes. Wash 3 time...
Embodiment 2
[0096] Example 2: Correlation between TLR3 expression and neutrophil infiltration in lung cancer paracancerous tissues and survival time of patients.
[0097] The expression of TLR3 and CD66b (reflecting the number of neutrophils) in the paracancerous tissues of 90 cases of lung cancer were analyzed by immunohistochemical method, and the expression of TLR3 and the infiltration of neutrophils in the paracancerous tissues of lung cancer were compared with those of patients. The correlation of survival time, the P value is calculated using the log-rank test in SPSS 17.0, and the results are as follows figure 2 A and figure 2 Shown in B.
[0098] figure 2 The result of A shows that the high expression of TLR3 is significantly related to the lower overall survival time of patients. figure 2 The results of B showed that more infiltration of neutrophils was significantly related to lower overall survival time of patients.
[0099] The above results indicated that the high exp...
Embodiment 3
[0100] Example 3: Regulation of TLR3 on tumor lung metastasis in mouse lung cancer and melanoma spontaneous lung metastasis models.
[0101] TLR3-deficient mice were purchased from Jackson Laboratory in the United States and raised in an SPF-grade environment. TLR3 gene-deficient mice and littermate wild-type control mice were obtained through crossbreeding. The experimental and control mice used in this study were female mice of C57BL / 6 background, aged 8-10 weeks.
[0102] Establishment of lung cancer and melanoma spontaneous lung metastasis models in mice: each mouse was subcutaneously injected with about 1×10 6 A lung cancer cell line LLC or melanoma B16 / F10 cells (the cells were purchased from PerkinElmer, USA). When the tumor grows to 1cm 2 When the size (18-20 days), dorsal tumors were resected. Lung metastases of mice were detected 40-45 days after inoculation.
[0103] Live imaging of lung tissue in vitro: LLC and B16 / F10 cells were infected with luciferase lentiv...
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