Chrono-biological genetherapy for bile duct cancer

癌细胞对抗肿瘤药物的敏感性是由癌细胞的增殖状态和细胞周期决定的，它是癌细胞的细胞周期的统一，药物治疗的时机和副作用少的癌症药物治疗是预期的。近年来，一种时钟基因的研究不断进展，分子生物学对各个时钟基因的研究也在不断进展，per1、per2、cry1和cry2、BMAL-1等作为主要的时钟基因备受关注，据说per2与癌变密切相关。在本研究期间，我们首先以最重要的per2的PAS结构域和CKI结构域为中心，验证了clock基因在各种癌细胞中的存在。这些细胞系检测了胆管癌细胞、TFK-1、HuCCT1、肝癌细胞、HepG2、Hep3B、HT-17、Li-7、HuH-7、PLC/PR/5、乳腺癌细胞、MCF-7、MDA-MB-231，并通过RT-PCR和序列检测了per2的存在。另一方面，制备用于基因治疗的腺病毒载体，用表达CDK抑制剂p27的腺病毒检测其对胆管癌的转导效率。CDK抑制剂p27与clock基因密切相关，控制细胞周期。细胞周期停止并诱导细胞凋亡至胆管癌（TFK-1）。Adp27。这一结果可以预期生物钟基因治疗的效果。

The susceptibility of the cancer cell to anti-neoplasm medicine is prescribed by the multiplication state of a cancer cell, and the cell cycle, it is uniting the cell cycle of a cancer cell, and the timing of medicine medication, and cancer medical treatment with few side effects is expected. Research of a clock gene is progressing recently, molecular biology-examination of each clock gene is progressing, per1, per2, cry1 and cry2, BMAL-1, etc. attract attention as main clock genes, and it is said that per2 is closely related with carcinogenesis.Existence of the clock gene in various cancer cells was checked centering on the PAS domain and CKI domain of per2 considered to be first the most important as a result in this research period. The cell lines examined bile duct cancer cells, TFK-1, the HuCCT1, liver cancer cell, HepG2, Hep3B, HT-17,Li-7,HuH-7,PLC/PR/5, breast cancer cells, MCF-7, and MDA-MB-231, and checked existence of per2 by RT-PCR and the sequence.On the other hand, preparing the adenoviral vector for genetherapy, Adenovirus expressing CDK inhibitor p27 was used for check efficiency of transduction for cholangiocarcinoma. CDK inhibitor p27 is closely related with clock gene as a control of cell cycle. The cell cycle stopped and induced apotosis to cholangiocarcinoma,TFK-1, using. Adp27. This result can expect the effect of the gene therapy by the clock gene.