MOLECULAR-BIOLOGICAL APPROACHES TO CARCINOGENESIS AND METASTASIS OF HEAD AND NECK CANCER AND THEIR APPLICATIONS TO IMMUNO-GENETIC THERAPY

头颈癌致癌和转移的分子生物学方法及其在免疫遗传学治疗中的应用

• 批准号: 13470362
• 负责人: YAMANAKA Noboru
• 金额: $ 5.18万
• 依托单位: WAKAYAMA MEDICAL 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 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470362/
• 关键词: HEAD AND NECK CANCER; BETA-CATENIN; APC; VEGF; TNP-470; SURVIVIN; 頭頚部癌; Β-カテニン; 癌抑制遺伝子; カドヘリン; 転移; 免疫組織染色

1) Beta-catenin accumulation and the mutation of the beta-catenin gene head and neck cancers were evaluated. Twelve out of 49 (24.5%) cases exhibited beta-catenin accumulation in our histochemical study. The 5 year survival rate was 0% in the beta-catenin accumulation group, compared to 50% in the non-accumulation group, (p<0.01). This finding strongly suggests that beta-catenin may play an important role in the carcinogenesis or progression of head and neck cancer. One of the 15 cases exhibited an APC missense mutation that led to the replacement of amino acids ; this case died in 12 months. Regarding the beta-catein mutation, non of the 31 samples exhibited a gene mutation in beta-catenin exon 3. Thus, the rate of APC and beta-catenin mutation in head and neck cancer may be very low.2) Vascular endothelial growth factor (VEGF) gene was transfected to the head and neck cancer cell line which lacked the VEGF gene and the effect of gene induction was examined in vitro and in vivo. The m … More axillary cancer cell line which was not expressing VEGF, OKK-LN was established and VEGF gene expressing vector was transfected to OKK-LN cell line and OKK-LN/pCIneo-VEGF which producing VEGF was established. This cell line extensively produced VEGF compared with the control cell line, OKK-LN/pCIneo transfected with pCIneo plasmid alone. Both cell lines were transplanted into 6 week-Balb/c nu/nu mice by subcutaneous injection to right lateral abdomen. Tumor volume of the VEGF-transfected group significantly increased in approximately three times compared with the control group after 35 days of transplantation. The survival of the VEGF-transfected group was shortened by 9 days on an average. The effect on the proliferation of VEGF-producing tumor by TNP-470, Fumagilin analogue, which nonspecifically inhibits angiogenesis was examined. Tumor volume of TNP-470 administered group was significantly reduced to one-third compared with non-administered group after 35 and 42 days of the transplantation. The survival of TNP-470 administered group was significantly prolonged. In conclusion, the production of VEGF may facilitate tumor proliferation, influence the survival, and contribute to the malignancy and the suppression of VEGF may be one of the promising strategies against head and neck cancers.3) We examined the expression of survivin and the two splicing variants in various cancer cells and normal adult tissues. Two splicing variants were detected in various types of cancer cells as well as survivin. By stimulating peripheral blood lymphocytes with survivin-pulsed antigen-presenting cells, CTLs were successfully induced in vitro from HLA-A24-positive cancer patients. These data suggest that survivin may be a potent T-cell epitope eliciting CTL response against survivin, which is expressed in many kinds of cancer cells. Less

1)β-连环蛋白积累和β-连环蛋白基因突变的头颈癌进行了评估。在我们的组织化学研究中，49例中有12例（24.5%）显示β-连环蛋白积聚。β-连环蛋白积聚组的5年生存率为0%，而非积聚组的5年生存率为50%（p<0.01）。这一发现有力地表明，β-连环蛋白可能在头颈癌的发生或进展中发挥重要作用。15例中有1例表现出APC错义突变，导致氨基酸替换;该病例在12个月内死亡。关于β-连环蛋白突变，31个样本中没有一个在β-连环蛋白外显子3中表现出基因突变。因此，APC和β-catenin在头颈癌中的突变率可能很低。2）将血管内皮生长因子（VEGF）基因转染到缺失VEGF基因的头颈癌细胞系中，并在体外和体内检测基因诱导的效果。m个 ...更多信息 建立不表达VEGF的腋窝癌细胞株OKK-LN，并将VEGF基因表达载体转染到OKK-LN细胞株中，建立产生VEGF的OKK-LN/pCIneo-VEGF。与对照细胞系（仅用pCIneo质粒转染的OKK-LN/pCIneo）相比，该细胞系广泛地产生VEGF。通过右腹部皮下注射将两种细胞系移植到6周-Balb/c nu/nu小鼠中。移植35天后，VEGF转染组的肿瘤体积与对照组相比显著增加了约3倍。VEGF转染组的生存期平均缩短了9天。研究了非特异性抑制血管生成的烟曲霉素类似物TNP-470对产生VEGF的肿瘤增殖的影响。在移植后35和42天，TNP-470给药组的肿瘤体积与未给药组相比显著减少至三分之一。TNP-470治疗组的生存期明显延长。总之，VEGF的产生可能促进肿瘤的增殖，影响生存，并有助于恶性肿瘤的发生，抑制VEGF可能是治疗头颈部肿瘤的有效策略之一。3）我们检测了Survivin及其两种剪接变异体在各种癌细胞和正常成人组织中的表达。在各种类型的癌细胞以及生存素中检测到两种剪接变体。通过用存活素脉冲的抗原呈递细胞刺激外周血淋巴细胞，成功地在体外从HLA-A24阳性的癌症患者中诱导CTL。这些数据表明，生存素可能是一个有效的T细胞表位引发针对生存素的CTL应答，生存素在多种癌细胞中表达。少

项目成果

竹井慎, 他: "頭頚部癌における予後因子としてのΒカテニンの過剰発現とAPC遺伝子変異の検討"日本耳鼻咽喉科学会誌. (印刷中). (2003)

Shin Takei 等人：“β-连环蛋白和 APC 基因突变作为头颈癌预后因素的检查”，日本耳鼻喉科学会杂志（2003 年出版）。

上野ゆみ他: "頭頸部腫瘍におけるVEGF発現に関する検討"耳鼻咽喉科免疫アレルギー. 19. 176-177 (2001)

Yumi Ueno 等：“头颈肿瘤中 VEGF 表达的研究”耳鼻喉科免疫学和过敏学 19. 176-177 (2001)。

Ueno Y, et al.: "Implication of Vascular Endothelial Growth Factor (VEGF) in Head and Neck Cancer"J Jp Immunol Allergol ORL. 19. 176-177 (2001)

Ueno Y 等人：“血管内皮生长因子 (VEGF) 在头颈癌中的意义”J Jp Nutritional Allergol ORL。

竹井慎, 他: "頭頚部癌におけるWintシグナル経路の検討"耳鼻咽喉科免疫アレルギー. 19. 174-175 (2001)

Shin Takei 等人：“头颈癌中 Wint 信号通路的检查”《耳鼻喉科免疫学和过敏》 19. 174-175 (2001)。

Hirohashi Y, et al.: "An, HLA-A24-restricted cytotoxic, T-lymphocyte epitope of a tumor-associated protein, survivin"Clin Cancer Res. 8(6). 1731-1739 (2002)

Hirohashi Y 等人：“肿瘤相关蛋白存活蛋白的 HLA-A24 限制性细胞毒性 T 淋巴细胞表位”Clin Cancer Res。