Molecular, biological studies on the mechanism of neck metastasis from nasopharyngeal carcinoma

鼻咽癌颈部转移机制的分子生物学研究

• 批准号: 13470358
• 负责人: FURUKAWA Mitsuru
• 金额: $ 7.55万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470358/
• 关键词: LMP1; MMP9; NF-κB; I-κB; Invasiveness; Metastasis; Gene targeted therapy; Aspirin; I-_κB; bFGF; VEGF; TGF-α; IL-8

Nasopharyngeal carcinoma(NPC), an epithelial tumor which is characterized by marked geographic and population differences in incidence, is found to be associated with Epstein-Barr virus(EBV) by serologic evidence, and the relationship was confirmed by the detection of EBVDNA and EB-encoded RNAs in NPC cells. While, NPC is highly metastatic carcinoma whose consistent associated with EBV has been established. Latent membrane protein 1(LMP1), an EBV membrane protein expressed in latent infection is considered to be the EBV oncoprotein. Matrix metalloproteinase 9(MMP9), one of the MMP families, degrades Type IV collagen, a major 4 component of extracellural matrix and is believed to be crucial for cancer invasion and metastasis. Although MMP9 is reported to be expressed in a variety of cancers, no reports concerning NPC have been published. We have shown that LMP1 induces MMP9 in vitro cell line, which suggests the possibility of mechanism in which LMP1 of EBV contributes to the metastasis … More and tumorgenesis of NPC by the induction of MMP9. Then the expression of LMP1 and MMP9 were immunohistochemically examined, and the relation of these proteins was statistically analyzed. We also analyzed the association of these proteins with clinical features. As results, both LMP1 and MMP9 proteins were predominantly immunolocalized in cancer nests. The expression of MMP9 showed a significant positive correlation with the expression of LMP1. Also, the expression of MMP9 correlated with lymphnode metastasis.We also demonstrated that LMP1 enhances MMP9 expression by activation of nuclear factor(NF)κB and activator protein(AP)-1. We therefore tested whether up-regulation of MMP9 by LMP1 could be correlated with enhanced invasiveness of tumor cells in vitro. Whether aspirin and sodium salicylate could reduce invasiveness and whether LMP1 could enhance MMP9 expression in tumors grown in nude mice were also tested. CS3A cells stably expressing LMP1 had increased expression of MMP9 and showed greater invasion through reconstituted basement membrane compared with vector-transfected C33A cells. Treatment with aspirin and sodium salicylate inhibited invasiveness of the LMP1-expressing C33A cells and suppressed both the LMP1-induced MMP9 expewssion in zymographic analyses and LMP1-induced MMP9 promoter activity in CAT reporter assays. The inhibitory effect of aspirin on NF-κB activity was attributable to the inhibition of I-κB kinase activity. Finally, tumors derived from vector-transfected C3SA cells stably expressing LMP1 grown in nude mice showed enhanced MMP9 levels compared with tumors derived from vector-trancfected C33A cells. This enhancement was inhibited by treatment of the mice with aspirin. These results suggest that aspirin may be able to suppress invasion and metastasis of EBV-associated tumors that express LMP1 by suppression of MMP-9. Less

鼻咽癌（Nasopharyngeal carcinoma，NPC）是一种具有明显地理和人群差异的上皮性肿瘤，血清学研究发现其与EB病毒（Epstein-Barr virus，EBV）相关，并通过检测NPC细胞中EB病毒DNA和EB病毒编码的RNA证实了这种关系。而鼻咽癌是一种高转移性肿瘤，其与EBV的相关性已被证实。潜伏膜蛋白1（LMP 1），在潜伏感染中表达的EBV膜蛋白，被认为是EBV癌蛋白。基质金属蛋白酶9（MMP 9）是MMP家族之一，可降解IV型胶原蛋白（细胞外基质的主要成分），被认为对癌症侵袭和转移至关重要。虽然据报道MMP 9在多种癌症中表达，但尚未发表关于NPC的报道。我们已经证明LMP 1在体外细胞系中诱导MMP 9，这表明EBV的LMP 1参与转移的机制的可能性。 ...更多信息 MMP 9诱导鼻咽癌的发生。采用免疫组化法检测LMP 1和MMP 9的表达，并进行相关性分析。我们还分析了这些蛋白与临床特征的关联。结果，LMP 1和MMP 9蛋白主要在癌巢中免疫定位。MMP 9与LMP 1的表达呈显著正相关。MMP 9的表达与淋巴结转移密切相关，LMP 1通过激活核因子（NF）κB和激活蛋白（AP）-1促进MMP 9的表达。因此，我们测试了LMP 1上调MMP 9是否与体外肿瘤细胞侵袭力增强相关。还测试了阿司匹林和水杨酸钠是否可以降低侵袭性，以及LMP 1是否可以增强裸鼠肿瘤中MMP 9的表达。稳定表达LMP 1的CS 3A细胞MMP 9的表达增加，并且与载体转染的C33 A细胞相比，表现出更大的通过重建基底膜的侵袭。用阿司匹林和水杨酸钠处理抑制表达LMP 1的C33 A细胞的侵袭性，并抑制酶谱分析中LMP 1诱导的MMP 9表达和CAT报告基因分析中LMP 1诱导的MMP 9启动子活性。阿司匹林对NF-κB活性的抑制作用可能与抑制I-κB激酶活性有关。最后，与来自载体转染的C33 A细胞的肿瘤相比，来自载体转染的C3 SA细胞的肿瘤在裸鼠中稳定表达LMP 1，显示出增强的MMP 9水平。这种增强被阿司匹林治疗的小鼠所抑制。这些结果表明，阿司匹林可能能够通过抑制MMP-9来抑制表达LMP 1的EBV相关肿瘤的侵袭和转移。少

项目成果

T.Horikawa: "Short Communication-induction of c-Met Proto-Oncogene by Epstein-Barr Virus Latent Membrane Protein-1 and Correlation with Cervical Lymph Node Metastasis of Nasopharyngeal Carcinoma"American Journal of Pathology. 159・1. 27-33 (2001)

T. Horikawa：“Epstein-Barr病毒潜伏膜蛋白1对c-Met原癌基因的短通讯诱导及其与鼻咽癌颈部淋巴结转移的相关性”美国病理学杂志159・1。 2001）

吉崎 智一: "上咽頭癌転移関連遺伝子発現と遺伝子治療の可能性-アスピリンは転移を抑制するか-"日本耳鼻咽喉科学会会報. 104. 791-795 (2001)

Tomokazu Yoshizaki：“鼻咽癌转移相关基因表达和基因治疗的可能性 - 阿司匹林会抑制转移吗？”日本耳鼻喉科学会通报 104. 791-795 (2001)。

吉崎 智一: "舌癌におけるMMP2活性化関連遺伝子発現と治療戦略"頭頸部腫瘍. 27・3. 659-662 (2001)

Tomokazu Yoshizaki：“舌癌中MMP2激活相关基因的表达和治疗策略”头颈肿瘤27・3（2001）。

H.Kinsen, H.Sato, M.Furukawa, T.Yoshizaki: "Modulation of Cell Growth and Matrix Metalloproteinase-2 Activation of Oral Squamous Cell Carcinoma as a Function of Culture Condition with Type 1 Collagen"Acta Otolaryngol. 123. 987-993 (2003)

H.Kinsen、H.Sato、M.Furukawa、T.Yoshizaki：“口腔鳞状细胞癌的细胞生长和基质金属蛋白酶-2 激活的调节作为 1 型胶原培养条件的函数”Acta Otolaryngol。

T.Yoshizaki, H.Sato, M.Furukawa: "Recent advances in the regulation of matrix metalloproteinase 2 activation : From basic research to clinical implication"(Review) Oncology Report. 9. 607-611 (2002)

T.Yoshizaki、H.Sato、M.Furukawa：“基质金属蛋白酶 2 激活调节的最新进展：从基础研究到临床意义”（综述）肿瘤学报告。