小鼠口腔癌不同时期淋巴结和骨髓的VEGF表达情况及其微环境对淋巴道转移的影响

目的 探讨小鼠口腔癌不同时期淋巴结和骨髓的血管内皮生长因子(VEGF)表达情况,及其微环境对口腔癌淋巴道转移的影响.方法 选入40只6周龄雄性健康Balb/c小鼠,将其随机分为A、B、C、D、E组,每组8只.采用4-硝基喹啉-1-氧化物(4-NQO)饮水法构建小鼠口腔癌淋巴道转移模型.其中A组为正常小鼠;B组小鼠舌部黏...

The expressions of VEGF in lymph nodes and bone marrow of mice with oral cancer at different stages and the influences of their microenvironments on lymphatic metastasis

Objective To explore the expressions of vascular endothelial growth factor(VEGF)in lymph nodes and bone marrow of mice with oral cancer at different stages,and the influences of their microenvironments on lymphatic metastasis of oral cancer.Methods Forty 6-week-old male healthy Balb/c mice were selected and randomly divided into groups A,B,C,D,and E,with 8 mice in each group.The 4-nitroquinoline-1-oxide(4-NQO)drinking water method was used to construct a mouse oral cancer lymphatic metastasis model.Among them,group A was normal mice;group B had no pathological changes in the tongue mucosas of the mice,but there were already 4-NQO drug effects;group C had abnormal proliferation in the tongue mucosas of the mice,but no cancer had formed;group D had developed tongue cancer of the mice,but no transfer had taken place;group E had developed oral cancer of the mice,accompanied by mandibular lymph node metastasis.Immunohistochemistry and liquid chip methods were used to detect the expressions of VEGF in lymph nodes and bone marrow of the mice in different groups and the results were compared.The effects of lymph node and bone marrow microenvironments on the expressions of VEGF in the human tongue squamous cell carcinoma cells(SAS cells)in groups A,B,C and D were observed.Results The results of VEGF immunohistochemical staining showed that VEGF expression was seen in the lymph nodes of the five groups,and the cytoplasmic staining was positive;however,the same bone marrow section under different visual fields was very different and difficult to analyze.The liquid chip method test results showed that for the lymph node tissue,the VEGF expression level of group A was significantly higher than that of group C and D(P<0.05),and the VEGF expression level of group E was significantly higher than that of group C(P<0.05),and there was no statistically significant difference between the other groups(P>0.05).For the bone marrow tissue,the VEGF expression levels of group A and group B were significantly higher than those of group C,group D and group E(P<0.05),and there was no statistically significant difference between the other groups(P>0.05).The expression levels of VEGF in the bone marrow tissue of the mice in the five groups were significantly higher than those in the lymph node tissue of the mice in the five groups(P<0.05).The results of co-cultivation experiments showed that in the microenvironments of lymph nodes and bone marrow in the periods of group A,group B and group C,the expression of VEGF in SAS cells showed an upward trend during the 24-72 h period.In the microenvironment of lymph nodes in the period of group D,the expression of VEGF in SAS cells showed a downward trend during the 24-72 h period,but in the microenvironment of bone marrow in the period of group D,the expression of VEGF in SAS cells showed an increased trend during the 24-72 h period.Conclusion Lymph node and bone marrow microenvironments may affect the expression of VEGF in disseminated tumor cells,so as to achieve the targeting of oral cancer lymphatic metastasis.