A study on the prevention of oral cancer metastasis by targeting a transcription factor NF-κB and protease

靶向转录因子NF-κB和蛋白酶预防口腔癌转移的研究

• 批准号: 11671993
• 负责人: IKEBE Tetsuro
• 金额: $ 1.98万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671993/
• 关键词: NF-κB; MMP; oral cancer; tumor invasion and metastasis

We examined a validity of NF-κB targeting therapy of oral cancer. Firstly, the involvement of matrix metalloproteinase-9 (MMP-9) in cancer invasion and metastasis was studied in the biopsy specimens of oral cancer patients. Secondly, in order to block the capability of oral cancer cells to produce MMP-9, we attempted to inhibit the activation of a transcription factor NF-κB through various methods.(1) We examined a gelatinolytic activity of MMP-9 in the biopsy specimens of 57 cases of oral squamous cell carcinoma by gelatin zymography. The same specimens were also stained immunohistochemically by anti-MMP-9 antibody. The gelatinolytic activity of MMP-9 in each case correlated with the immunohistochemical staining degree of MMP-9. The gelatin zymographic analysis of 57 cases revealed that the gelatinolytic activity of MMP-9 in biopsy specimens statistically correlated with histopathological invasiveness of oral cancer, suggesting that MMP-9 expression contributes to the invasion and met … More astasis of oral cancer.(2) We found that the MMP-9 expression of cultured oral squamous cell carcinoma cell lines was inhibited by dexamethasone (DEX) and interleukin-4 (IL-4). DEX and IL-4 inhibited the conversion from MMP-9 precursor to active form of MMP-9 by inhibiting the gene expression of urokinase type plasminogen activator (uPA). The activation of a transcription factor NF-κB, which can activate gene expressions of MMP-9 and uPA, was also inhibited by DEX and IL-4. Therefore, DEX and IL-4 are likely to target NF-κB to inhibit the gene expression and activation of MMP-9.(3) Early-passage normal human fibroblasts can express normal size of NF-κB while the molecular size of NF-κB of late-passage fibroblasts was reduced to 70-80 kD.The senescence of normal cells may induce the intracellular NF-κB-degrading protease(s). In contrast, tumor cells can express normal NF-κB molecule stably evev after 100 passage.(4) The expression vectors of superreppressor IκBα and dominant negative IκBα kinases (IKK kinases) have been formed. We are now transtecting the expression vectors into oral cancer cells and examining the effects of these vectors on MMP-9 expression.In conclusion, NF-κB-targeting therapy may be useful to prevent invasion and metastasis of oral cancer. Less

我们研究了NF-κB靶向治疗口腔癌的有效性。本研究首先在口腔癌患者的活检标本中研究了基质金属蛋白酶-9（MMP-9）在口腔癌侵袭和转移中的作用。其次，为了阻断口腔癌细胞产生MMP-9的能力，我们尝试通过各种方法抑制转录因子NF-κB的活化。(1)我们用明胶酶谱法检测了57例口腔鳞状细胞癌活检标本中MMP-9的明胶酶活性。同样的标本也用抗MMP-9抗体进行免疫组化染色。MMP-9的明胶溶解活性与MMP-9的免疫组化染色程度相关。57例口腔癌组织中MMP-9的明胶酶谱分析显示，MMP-9的明胶酶活性与口腔癌的组织病理学侵袭性相关，提示MMP-9的表达与口腔癌的侵袭性有关， ...更多信息 口腔癌的复发(2)我们发现地塞米松（DEX）和白细胞介素-4（IL-4）可抑制体外培养的口腔鳞癌细胞系MMP-9的表达。DEX和IL-4通过抑制尿激酶型纤溶酶原激活物（uPA）基因表达，抑制MMP-9前体向活性型的转化。DEX和IL-4也可抑制转录因子NF-κB的活化，NF-κB可激活MMP-9和uPA的基因表达。因此，DEX和IL-4可能靶向NF-κB，抑制MMP-9的基因表达和活化。(3)早期传代的正常人成纤维细胞可表达正常大小的NF-κB，而晚期传代的成纤维细胞NF-κB的分子量减少到70-80 kD，正常细胞的衰老可能诱导细胞内NF-κ B降解蛋白酶的产生。而肿瘤细胞在传代100代后仍能稳定表达正常NF-κB分子。(4)超表达IκBα和显性负性IκBα激酶（IKK kinases）的表达载体已经构建。本研究将NF-κ B基因转染口腔癌细胞，观察其对口腔癌细胞MMP-9表达的影响，为NF-κ B靶向治疗口腔癌提供实验依据。少

项目成果

Ikebe,T.,Shinohara,M.,Takeuchi,H.,Beppu,M.,Kurahara,S.,Nakamura,S.その他: "Gelatinolytic activity of matrix metalloproteinase in tumor tissues correlates with the invasiveness of oral cancer"Clinical & Experimental Metastasis. 17. 315-323 (1999)

Ikebe, T.、Shinohara, M.、Takeuchi, H.、Beppu, M.、Kurahara, S.、Nakamura, S. 等人：“肿瘤组织中基质金属蛋白酶的明胶分解活性与口腔癌的侵袭性相关”临床与实验转移。17. 315-323 (1999)