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

口腔鳞状细胞癌是口腔颌面部最常见的恶性肿瘤，由于肿瘤的表面性，口腔鳞癌具有肉眼可识别的特点。首先，我们检测了E1AF在正常组织和肿瘤组织中的表达。E1AF是一种非癌基因家族转录因子，我们已经注意到E1AF可以上调多种基质金属蛋白酶(MMPs)基因的启动子活性，并表明口腔鳞癌来源的细胞系的侵袭潜能与E1AF和MMPs的表达有关。我们分析了E1AF在非小细胞肺癌(NSCLC)细胞系和切除肿瘤中的表达。17个非小细胞肺癌细胞系中有15个细胞株和19个肿瘤细胞系中有12个细胞表达E1AF基因，而正常肺组织和肿瘤内伴发的正常细胞均不表达E1AF基因。在口腔癌和正常组织中也得到了类似的结果。逆转录聚合酶链式反应检测E1AF的表达。27例患者中有15例检测到E1AF，正常组织中未检测到E1AF。域名。目前，只有BH3蛋白被认为是细胞凋亡信号的始发者。我们的结果表明，BnipS通过TM结构域将caspase募集到线粒体膜上，从而诱导细胞凋亡。将鳞状细胞癌细胞系SiHA移植到裸鼠体内。瘤内应用Ad-F-Bik，每周两次。应用Ad-F-Bik抑制肿瘤生长，诱导肿瘤细胞凋亡，结果表明，用腺病毒载体靶向肿瘤特异性表达分子的基因治疗是有效的。

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.