Gene therapy for oral squamous cell carcinoma targeting tumor specific promoter activation

靶向肿瘤特异性启动子激活的口腔鳞状细胞癌基因治疗

• 批准号: 12671834
• 负责人: SHINDOH Masanobu
• 金额: $ 2.24万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671834/
• 关键词: E1AF; adenvirus vector; promoter activity; survivin; 転写調節領域

Oral squamous cell carcinoma is the mostcommon malignant tumor in oral and maxillofacial region, and has a property that oral carcinoma is able to be recognized macroscopy because of the superficial occurrence of tumors. This characteristics contribute to the gene targeting therapy.First, we examined E1AF expression in normal and tumor tissues. E1AF is anete-oncogene family transcription factor, and we have noted that E1AF can upregulate promoter activities of several matrix metalloproteinase (MMP) genes and showed that invasive potentials of oral squamous cell carcinoma-derived cell lines are correlated with expression of E1AF and MMPs. We analyzed the E1AF expression in cell lines and resected tumors of non-small-cell lung cancers (NSCLC). Fifteen out of 17 cell lines of NSCLC and 12 of 19 tumors expressed E1AF mRNA while normal lung tissue and concomitant normal cells within tumors did not express E1AF mRNA. Similar results were obtained in oral carcinoma and normal tissues. RT-PCR was utilizedto examine the expression of E1AF. Among the 27 patients, E1AF was detected in 15 cases and was not detected in normal tissues.Bnip3 is a pro-apoptotic molecule that contains BH3 and trans-membrane ? domains. BH3 only proteins are now thought to be the initiator of apoptotic signals. Our results suggest that BnipS induces apoptosis by recruiting caspases onto mitochondrial membrane through TM domain.Bik, a BH3 only molecule, expressing adenovirus vector driven by E1AF promoter was reconstructed (Ad-F-Bik). SiHa, a squamous cell carcinoma cell line was transplanted into nude mice. Ad-F-Bik was applied in vivo tumors twice a week. Tumor growth was inhibited by application of Ad-F-Bik, and apoptosis of tumor cells was induced.These results indicate that gene therapy targeting tumor specific expressing molecule is effective using adenovirus vector.

口腔鳞状细胞癌是口腔颌面部最常见的恶性肿瘤，具有肿瘤发生表浅、肉眼即可识别的特点。首先，我们检测了E1 AF在正常组织和肿瘤组织中的表达。E1 AF是一种癌基因家族转录因子，我们发现E1 AF可以上调多种基质金属蛋白酶（MMP）基因的启动子活性，并发现口腔鳞癌细胞系的侵袭能力与E1 AF和MMP的表达相关。我们分析了E1 AF在非小细胞肺癌（NSCLC）细胞系和切除肿瘤中的表达。15/17的NSCLC细胞系和12/19的肿瘤表达E1 AF mRNA，而正常肺组织和肿瘤内伴随的正常细胞不表达E1 AF mRNA。在口腔癌和正常组织中获得了类似的结果。RT-PCR检测E1 AF基因的表达。Bnip 3是一种促凋亡分子，含有BH 3和跨膜调节因子（transmembrane？域.现在认为只有BH 3蛋白质是凋亡信号的启动子。结果表明BnipS通过TM结构域将caspase募集到线粒体膜上而诱导细胞凋亡。将人鳞状细胞癌细胞株SiHa移植到裸鼠体内。Ad-F-Bik每周两次应用于体内肿瘤。Ad-F-Bik可抑制肿瘤生长，诱导肿瘤细胞凋亡，表明以肿瘤特异性表达分子为靶点的腺病毒载体基因治疗是有效的。

项目成果

Yasuda M, Yamazaki K, Hamahira S, Kawasaki T, Nakamura M, Shindoh M: "Functional dissection of BH3 only protein BNIP3."Tumor Res. 36. 23-28 (2001)

Yasuda M、Yamazaki K、Hamahira S、Kawasaki T、Nakamura M、Shindoh M：“仅 BH3 蛋白 BNIP3 的功能解剖。”肿瘤研究。

Hiroumi H, Dosaka-Akita H, Shindoh, M et al.: "Expression of E1AF an ets-related transcription factor in human non-small cell lung cancer"Int J Cancer. 93. 786-791 (2001)

Hiroumi H、Dosaka-Akita H、Shindoh、M 等人：“E1AF 是人类非小细胞肺癌中 ets 相关转录因子的表达”Int J Cancer。

Yasuda M, Yamazaki K, Shindoh M et al.: "Functional dissection of BH3 only protein BNIP3"Tumor Res. 36. 23-28 (2001)

Yasuda M、Yamazaki K、Shindoh M 等人：“BH3 唯一蛋白 BNIP3 的功能解剖”肿瘤研究。

Hiroumi H, Dosaka-Akita H, Yoshida K, Shindoh M, Ohbuchi T, Fujinaga K, Harada T, Hommura F, Ogura S, Kawakami Y: "Expression of E1AF an ets-related transcription factor in human non-small cell lung cancers:Its relevance in cell motility and invasion."Int

Hiroumi H、Dosaka-Akita H、Yoshida K、Shindoh M、Ohbuchi T、Fujinaga K、Harada T、Hommura F、Ogura S、Kawakami Y：“E1AF（一种 ets 相关转录因子）在人类非小细胞肺癌中的表达