Shuyi Yu, Feng Wang, Jing Tang, Hongzhang Huang, Haiyan Wang, Pingping Ma, Liwu Zheng, Zwahlen Roger A, Hongyu Yang
Shenzhen PKU-HKUST Medical Center, Shenzhen, China.
Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011 Nov;112(5):616-25. doi: 10.1016/j.tripleo.2011.02.053.
In this study, the hypothesis that hBD-3 is upregulated by LPS via epidermal growth factor receptor (EGFR) signaling pathways to enhance metastasis in oral squamous cell carcinoma (OSCC) was tested.
hBD-3 expression in human tissue specimens was evaluated by RT-qPCR and immunohistochemical staining. The presence of hBD-3 peptide in the culture supernatants of each type of treated cells was evaluated by enzyme-linked immunosorbent assay. The chemotaxis response to LPS or hBD-3 protein of SCC-25 cells or siRNA-hBD-3 transfected cells were also measured by chemotaxis assay. Paired, 2-tailed Student t test and analysis of variance was used to assess the statistical significance between 2 groups or many groups.
hBD-3 is highly expressed and associated with lymphatic invasion of OSCC. hBD-3 expression and EGFR phosphorylation were markedly upregulated when SCC-25 cells were treated with LPS. When SCC-25 cells were preincubated with EGFR inhibitor or TLR4-neutralizing Ab before LPS stimulation, a decrease in the expression of hBD-3 was observed. hBD-3 markedly enhanced cancer metastasis, and the chemotaxis response to LPS of SCC-25 cells was partly blocked by siRNA target hBD-3.
These findings indicate that hBD-3 is upregulated by LPS via EGFR signaling pathways to enhance lymphatic invasion of OSCC.
在本研究中，检测了如下假说，即脂多糖（LPS）通过表皮生长因子受体（EGFR）信号通路上调人β-防御素3（hBD-3），从而增强口腔鳞状细胞癌（OSCC）的转移。
通过逆转录定量聚合酶链反应（RT-qPCR）和免疫组化染色评估hBD-3在人体组织标本中的表达。通过酶联免疫吸附测定评估每种处理细胞的培养上清液中hBD-3肽的存在情况。还通过趋化性测定测量SCC-25细胞或转染了小干扰RNA-hBD-3（siRNA-hBD-3）的细胞对LPS或hBD-3蛋白的趋化反应。采用配对双尾学生t检验和方差分析评估两组或多组之间的统计学显著性。
hBD-3高表达且与OSCC的淋巴浸润相关。用LPS处理SCC-25细胞时，hBD-3表达和EGFR磷酸化显著上调。在LPS刺激前，当SCC-25细胞与EGFR抑制剂或Toll样受体4（TLR4）中和抗体预孵育时，观察到hBD-3表达下降。hBD-3显著增强癌症转移，并且针对hBD-3的siRNA部分阻断了SCC-25细胞对LPS的趋化反应。
这些发现表明，LPS通过EGFR信号通路上调hBD-3，从而增强OSCC的淋巴浸润。
