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  

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

作　　者：路莹 程立 颜启璋 陈念 李晶[2] 郭梦竹 于大海[2] LU Ying;CHENG Li;YAN Qi-zhang(College of Stomatology,Guangxi Medical University,Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction,Guangxi Clinical Research Center for Craniofacial Deformity,Guangxi Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment(Key Laboratory of Guangxi Higher Education),Nanning 530021,China)

机构地区：[1]广西医科大学口腔医学院,广西口腔颌面修复与重建研究自治区级重点实验室,广西颅颌面畸形临床医学研究中心,颌面外科疾病诊治研究重点实验室(广西高校重点实验室),南宁530021 [2]广西医科大学第一附属医院口腔科,南宁530021

出　　处：《中国临床新医学》2021年第1期45-50,共6页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：广西自然科学基金重点项目(编号:2016GXNSFDA380002,2018GXNSFAA050088);国家自然科学基金项目(编号:81360407)。

摘　　要：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组小鼠舌部黏膜尚未出现病理改变,但已经存在4-NQO药物影响;C组小鼠舌部黏膜已经出现异常增生表现,但未形成癌症;D组小鼠已经形成舌癌,但尚未发生转移;E组小鼠口腔癌已经形成,并伴有下颌淋巴结转移。采用免疫组化法及液相芯片法检测不同组小鼠淋巴结和骨髓VEGF的表达情况,并进行比较。观察A、B、C、D组的淋巴结、骨髓微环境对人舌鳞状细胞癌细胞系(SAS细胞)VEGF表达的影响。结果VEGF免疫组化染色结果显示5组淋巴结均可见VEGF表达,胞浆染色阳性;但不同视野下,同一骨髓切片差异较大,难以进行分析。液相芯片法检测结果显示,对于淋巴结组织,A组VEGF表达水平显著高于C组和D组(P<0.05),E组显著高于C组(P<0.05),其余组间比较差异无统计学意义(P>0.05)。对于骨髓组织,A组和B组VEGF表达水平显著高于C组、D组和E组(P<0.05),其余组间比较差异无统计学意义(P>0.05)。5组小鼠骨髓组织的VEGF表达水平均显著高于淋巴结组织(P<0.05)。共培养实验结果显示,在A组、B组和C组时期的淋巴结和骨髓微环境中,SAS细胞在24~72 h时段内的VEGF表达呈上升趋势。而在D组时期淋巴结微环境中,SAS细胞在24~72 h时段内的VEGF的表达呈下降趋势,但在D组时期骨髓微环境中,SAS细胞在24~72 h时段内的VEGF的表达呈上升趋势。结论淋巴结及骨髓微环境可能会影响播散肿瘤细胞VEGF的表达,从而实现口腔癌淋巴道转移靶向性。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,g

关 键 词：口腔癌 血管内皮生长因子 播散肿瘤细胞 淋巴道转移 微环境 

分 类 号：R780.2[医药卫生—口腔医学]