Wnt1在口腔鳞癌中的表达及其与淋巴结转移关系的研究

目的 :研究Wnt信号传导通路中Wnt1蛋白在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达水平,初步探讨Wnt1在OSCC淋巴结转移中的作用。方法:采用免疫组织化学方法,检测60例OSCC中Wnt1的表达情况;采用Western blot法检测5对OSCC临床标本和配对癌周组织中Wnt1的表达;通过实时定量PCR法和Western blot法分别检测高、低转移潜能的HN-12和HN-4口腔鳞癌细胞系中Wnt1基因和蛋白的表达水平;采用生长曲线、划痕实验和Transwell法,分别检测重组蛋白Wnt1对2种细胞的生长情况、迁移能力和侵袭能力的影响。结果:24例淋巴结转移的OSCC中高表达Wnt1,16例未有淋巴结转移的OSCC中低表达Wnt1,Wnt1的表达在两组之间差异有统计学意义(P<0.01);高转移细胞系HN-12较低转移细胞系HN-4在m RNA水平和蛋白水平上Wnt1表达显著增高;重组蛋白Wnt1,能够提高肿瘤细胞的生长、迁移和侵袭能力。结论:Wnt信号通路中的关键分子Wnt1在OSCC中有表达,且与淋巴结转移相关,为进一步探讨Wnt信号通路影响肿瘤转移的分子机制奠定基础。

Wnt1 Correlates with Lymph Node Metastasis in OSCC

Objective: To study whether the expression of Wnt1 in patients with oral squamous cell carcinoma (OSCC) were correlated to lymph node metastasis. Methods: Immunohistochemistry was used to detect Wnt1 expression in OSCC tissues of 60 patients. Western blot was used to detect Wnt1 expression in 5 pairs of OSCC tissues and adjacent normal mucosa tissues . Real-time PCR and Western blot were used to detect Wnt1 expression on mRNA and protein levels re-spectively in HN-12 cells, which was highly metastatic compared with HN-4 cells. Growth curve, scratch test and tran-swell assay were used to detect the growth, migration and invasion ability of HN-12 and HN-4 cells which were treated with or without Wnt1 recombinant protein. Results: Wnt1 protein was up-regulated in 24 cases of OSCC with lymph node metastasis compared to 16 cases without lymph node metastasis (P<0.01). Moreover, the expression of Wnt1 on mRNA and protein levels was both much higher in HN-12 cells than HN-4 cells. Lastly, Wnt1 could notably enhanced cell growth, migration and invasion. Conclusion: The high expression of the key member Wnt1 of Wnt signal pathway was re-lated to lymph node metastasis of OSCC.