Oncolytic virus therapy for biliary and pancreas cancer using a replication-competent herpes simplex virus mutant and proapoptotic reagent

1)溶瘤病毒治疗胆囊癌的抗肿瘤疗效胆囊癌是一种一旦发生转移就极难治愈的疾病。在本文中，我们探讨了G207（一种溶瘤性、具有复制能力的单纯疱疹病毒1型突变体）作为胆囊癌治疗新手段的潜力。4例人、1例仓鼠胆囊癌细胞株在感染G207后72小时内几乎全部细胞死亡，MOI为0.25 ~ 2.5。对G207细胞病变活性的敏感性与lacZ表达的感染效率相关。在已建立的皮下KIGB-5肿瘤的具有免疫功能的仓鼠中，瘤内接种G207 （1 × 107 pfu）可显著抑制肿瘤生长并延长生存期。重复接种（3次，间隔4天）的效果明显高于单次接种。在双侧皮下KIGB-5肿瘤的仓鼠中，G207单独接种1…多个肿瘤可引起未接种肿瘤和接种肿瘤的消退或生长减少。然而，在胸腺小鼠中，接种肿瘤的抗肿瘤作用大大降低，而在远端肿瘤中完全消失，这表明T细胞介导的免疫应答对G207的局部和全身抗肿瘤作用都有很大贡献。这些结果提示G207可能作为胆囊癌治疗的新策略。2)溶瘤病毒与促凋亡因子对胰腺癌的联合作用促凋亡试剂Tetrocarcin A对G207体外对胰腺癌细胞的溶瘤作用没有增强作用。放疗和化疗药物联合使用没有增加溶瘤病毒活性。提示胰腺癌对促凋亡试剂具有抗性，或溶瘤病毒的细胞病变作用不依赖于细胞凋亡。3)溶瘤病毒对腹膜播散的治疗作用我们还阐明了溶瘤病毒对胆囊癌、结肠癌和胃癌腹膜播散的治疗作用，与对照组相比，G207使显微镜下腹膜播散动物的生存时间延长，部分动物治愈。另一方面，肉眼传播的动物虽然存活时间也比对照组延长，但没有治愈。以上数据已在分子治疗杂志2001年第3(4)期和外科治疗杂志2002年第87(3)期进行了总结和发表。少

1) Antitumor efficacy of oncolytic virus therapy for gallbladder cancerGallbladder cancer is an extremely difficult disease to cure once metastases occur. In this paper, we explored the potential of G207, an oncolytic, replication-competent herpes simplex virus type 1 mutant, as a new therapeutic means for gallbladder cancer. Gallbaladder carcinoma cell lines (4 human and 1 hamster) showed nearly total cell killing within 72 hours of G207 infection at a MOI of 0.25 to 2.5 in vitro. The susceptibility to G207 cytopathic activity correlated with the infection efficiency demonstrated by lacZ expression. Intraneoplastic inoculation of G207 (1 x 107 pfu) in immunocompetent hamsters bearing established subcutaneous KIGB-5 tumors caused a significant inhibition of tumor growth and prolongation of survival. Repeated inoculations (3 times with 4-day intervals) were significantly more effications than a single inoculation. In hamsters with bilateral subcutaneous KIGB-5 tumors, inoculation of one … More tumor alone with G207 caused regression or growth reduction of noninoculated tumors as well as inoculated tumors. In athymic mice, however, the antitumor effect was largely reduced in inoculated tumors and completely abolished in remote tumors, suggesting large contribution of T cell-mediated immune responses to both local and systemic antitumor effect of G207. These results indicate that G207 may be useful as a new strategy for gallbladder cancer treatment.2) Combination effect of oncolytic virus and proapoptotic factor for pancreas cancerA proapoptotic reagent, Tetrocarcin A, did not enhance the oncolytic effect of G207 for pancreas cancer cells in vitro. Combination of irradiation and chemotherapeutic reagents did not increase the oncolytic viral activity. These results suggest that pancreas cancer has a resistance to the proapoptotic reagent or the cytopathic effect of oncolytic virus does not depend on the apoptosis.3) Therapeutic efficacy of oncolytic virus for peritoneal disseminationWe also elucidated the therapeutic efficacy of oncolytic virus on peritoneal dissemination of gallbladder, colonic and gastric cancer, G207 prolonged the survival of animals bearing a microscopic peritoneal dissemination as compared with the controls, and some of the animals were cured. On the other hand, the animals harboring macroscopic dissemination were not cured although their survivals were also elongated than the control group.These data above have been summarized and published in Molechular Therapy 3(4), 2001 and Surgical Therapy 87 (3), 2002. Less