Lee Kyuhyun, Yun Sung-Tae, Kim Young-Gun, Yoon Yeup, Jo Eui-Cheol
Gene Therapy Laboratory, Biomolecular Engineering Division, MOGAM Biotechnology Research Institute, Yongin, Kyonggi-Do, Republic of Korea.
Hepatology. 2006 May;43(5):1063-73. doi: 10.1002/hep.21149.
Hepatocellular carcinoma (HCC) constitutes more than 90% of all primary liver cancers. HCC is a hypervascular tumor that develops from dedifferentiation of small avascular HCC and is therefore a good target for anti-angiogenic gene therapy. Recent studies have identified apolipoprotein(a) [apo(a)] kringles LK68 and LK8 (LKs) as having a potential antiangiogenic and anti-tumor activity, and the current study evaluates the therapeutic potential of gene therapy with recombinant adeno-associated virus carrying genes encoding LKs (rAAV-LK) in the treatment of hypervascular HCC. We generated rAAV-LK to obtain persistent transgene expression in vivo, which is essential for anti-angiogenic therapy. The rAAV-produced LKs substantially inhibited proliferation and migration of human umbilical vein endothelial cells (HUVECs) in vitro, validating their anti-angiogenic potential. Intramuscular administration of rAAV-LK gave 60% to 84% suppression (P < .05) of tumor growth in mice bearing subcutaneously transplanted HCC derived from Huh-7 and Hep3B cells, respectively. Histological and immunohistochemical analyses of HCC tumor sections showed that a single administration of rAAV-LK gave rise to persistent expression of LKs that inhibited tumor angiogenesis and triggered tumor apoptosis, and, thus, significantly suppressed tumor growth. The administration of rAAV-LK provided a significant survival benefit (P < .05), and 3 of 10 rAAV-LK-treated mice were still alive without visible tumors and without clinical symptoms 188 days after treatment. In conclusion, rAAV-LK is a potential candidate for anti-angiogenic gene therapy in the treatment of HCC.
肝细胞癌（HCC）占所有原发性肝癌的90%以上。HCC是一种血管丰富的肿瘤，由无血管的小HCC去分化发展而来，因此是抗血管生成基因治疗的良好靶点。最近的研究已确定载脂蛋白(a)[apo(a)] kringles LK68和LK8（LKs）具有潜在的抗血管生成和抗肿瘤活性，并且当前研究评估了携带编码LKs基因的重组腺相关病毒（rAAV-LK）基因治疗在治疗血管丰富的HCC中的治疗潜力。我们生成rAAV-LK以在体内获得持续的转基因表达，这对抗血管生成治疗至关重要。rAAV产生的LKs在体外显著抑制人脐静脉内皮细胞（HUVECs）的增殖和迁移，证实了它们的抗血管生成潜力。分别对皮下移植有源自Huh-7和Hep3B细胞的HCC的小鼠进行肌肉注射rAAV-LK，可使肿瘤生长抑制60%至84%（P <.05）。对HCC肿瘤切片进行组织学和免疫组化分析表明，单次注射rAAV-LK可导致LKs持续表达，抑制肿瘤血管生成并引发肿瘤凋亡，从而显著抑制肿瘤生长。注射rAAV-LK带来了显著的生存益处（P <.05），并且在治疗188天后，10只接受rAAV-LK治疗的小鼠中有3只仍然存活，没有可见肿瘤且没有临床症状。总之，rAAV-LK是HCC抗血管生成基因治疗的潜在候选者。
