【作者】 李刚；

【导师】 吴孟超； 张柏和； 卫立幸；

【作者基本信息】 第二军医大学 ， 外科学（肝外）， 2001， 硕士

【摘要】 肿瘤是目前危害人类健康的主要杀手之一。肝癌则被称为癌中之王，死亡率很高。目前肝癌的治疗仍以手术为主，辅以放化疗。由于手术和放化疗对机体的免疫系统，造血系统均有损害，以致疗效难以令人满意。近年来，基因治疗已成为恶性肿瘤治疗的研究热点，但其临床疗效并不令人满意，尚不能替代现行的常规治疗手段。但研究表明基因治疗具有增强机体免疫、提高放化疗敏感性、减少肿瘤的复发和转移等功效。因此，如何有效地将基因治疗和现行的常规肿瘤治疗方法有机的结合起来，互相补充，成为抗肿瘤研究的重要方向。 FL是近期发现的一种细胞因子，具有增强机体免疫和刺激造血的双重作用，我们希望FL和化疗联合应用，能克服化疗对机体免疫系统和造血系统的毒副作用，同时增强化疗的抗肿瘤效果。我们进行了如下实验：（1）MTT法检测Hepal-6肝癌细胞株对四种常用化疗药的敏感性，选择合适的化疗药；（2）携带鼠FLT3配体重组腺病毒的扩增、纯化、滴度测定和体外感染肝癌细胞；（3）化疗药和FLT3配体重组腺病毒联合治疗小鼠肝癌。 实验结果表明：（1）鼠Hepal-6肝癌细胞株对5-氟脲嘧啶和阿霉素相对敏感（2）FLT3配体重组腺病毒和阿霉素联合治疗具有协同作用，小鼠肝癌细胞生长抑制率由（单独应用阿霉素的）27.5％提高到56.5％，长期生存率由50％提高到86.4％；（3）FLT3配体重组腺病毒能大大降低阿霉素对造血系统的损害程度，化疗所致外周血白细胞下降的最低值由1.0×10~9/L提高到2.2×10~9/L，骨髓CFU-GM克隆数下降的最低值由4.3提高到12，并使其提前恢复。 国内对FLT3配体的研究主要集中在造血干细胞的体外扩增上， 中文摘要 对其抗肿瘤的作用研究较少：国外也未见应用F＊＊3配体重组腺病毒 对肝癌进行基因治疗的报道，尤其是应用FLT3配体重组腺病毒和化 疗联合应用。而且我们采用 F LT 3配体重组腺病毒肿瘤局部注射和全 身化疗相结合的方法，更符合临床治疗思路，并且取得了预期的实 验结果。希望我们的实验能为肝癌的临床治疗提供新思路，并为FLT3 配体应用孙＄床提供理论依据。 下一步的研究方向：（l）F LT 3 配体重组腺病毒和化疗药抗肿瘤 作用协同的机制：（二）＊＊＊3 配体重组腺病毒和化疗药抗肿瘤的中远 期效果；（3）改进腺病毒载体，降低载体本身的毒副作用：（4）FL＊3 配体和放疗相结合抗肿瘤治疗：（5）多基因联合治疗。更多还原

【Abstract】 At present cancer is one of the most dangerous disease and liver cancer is called 搕he king of the cancer? whose death rate is very high. Now the treatment of cancer is based on operation plus chemotherapy or radiation therapy. Since these methods are harmful to immune system and hematopoietic system, their curative effect cannot be satisfied. So recent years gene therapy becomes a hot spot of the field, but its clinical effect is not pleased . At the moment , gene therapy still cannot replace the conventional methods. However, research indicates that gene therapy can strengthen immunity, improve chemosensitivity, decrease the relapse and metastasis, so how to combine the gene therapy with conventional methods effectively becomes the main point of cancer research.The flt3-ligand (FL)is a recently identified cytokine ,whice can stimulate the proliferation of hematopoietic progenitors and enhance immune system. We hope that combination of FL and chemotherapy can relieve chemical drugs side effects on immune system and hematopoietic system , in mean time enhance its anti-tumor effect ,so we designed the followed experiments:1.Tested the chemo-sensitivity to anti-tumor agents in mice hepatocelluar carcinoma using a hepal-6 liver cancer cell line;2.Cloned the murine FL and constructed the recombinant adenoviral vector, paving the way for further cancer immunotherapy;3.Treated hepatocarcinoma in mice using murine FL recombinant adenovirus combined with chemotherapy.The results show:1.Adriamycine (ADM) and 5-fluorouracili (5-FU) are more effective to inhibit the growth of hepal-6 liver cancer cells;2.FL combined with ADM could significantly inhibit the growth of the liver cancer , make the rats survive long;3.FL could relieve ADM toxic effects on hematopoietic system ,make it recovery earlier.3-Domestic research on FL is focus on expansion of human cord blood cells in vitro ,not on its anti-tumor effects .Other countries have no reports on FL recombinant adenovirus activity against hepatoma , esp. on anti-tumor activity of FL combined with chemotherapy .We hope that these experiments can offer a new way for clinical cancer treatment and offer a theory for FL used in clinical treatment更多还原