Combined gene therapy with oncolytic adenovirus and NK-4 against tumor growth and invasion in hepatocellular carcinoma

过去用于治疗肝细胞癌（HCC）患者的许多方法不太令人满意。越来越多的证据表明，肝细胞生长因子（HGF）的拮抗剂NK 4抑制癌细胞的侵袭和转移，并且使用具有复制能力的腺病毒的基因治疗是癌症的有效治疗。肝癌，其中约70%可以产生甲胎蛋白（AFP），被认为是一种候选疾病的发展，一种新的治疗使用不同的基因和不同的载体。为了开发有效的抗癌治疗的合理基础，使用AFP启动子驱动的E1 B-55 k基因缺陷型腺病毒（AdAFP/Rep）和表达NK 4的腺病毒载体（AdCMV.NK4，由T.大坂大学的中村教授和T. Nukiwa，Tohoku University）靶向产生AFP的HCC细胞对肿瘤生长和侵袭的影响。AdAFP/Rep能够裂解所有的 ...更多信息 e产生AFP的HCC细胞系（HuH 7，HepG 2），并且不裂解原代培养物中的正常人肝细胞。NK 4对AFP阳性的肝癌细胞增殖有抑制作用，但对正常肝细胞无抑制作用。AdCMV.NK4处理可抑制产生AFP的HCC细胞通过Matrigel基底膜成分侵入胶原凝胶。用AdAFP/Rep或AdCMV.NK4治疗裸鼠移植的产生AFP的HCC细胞导致肿瘤生长的抑制。AdCMV.NK4处理降低HCC细胞和皮下移植瘤中血管内皮生长因子（VEGF）和CD 31以及基质金属蛋白酶（MMP）-2和MMP-9的mRNA表达。虽然AdCMV. NK 4单独使用并不比AdAFP/Rep具有更强的促凋亡作用，但AdAFP/Rep和AdCMV. NK 4联合使用可诱导细胞和肿瘤的协同促凋亡作用，并且抑制不可见HCC的生长和转移。这些结果清楚地表明，AdAFP/Rep和AdCMV.NK4的组合可以抑制实验动物HCC细胞的肿瘤生长、侵袭和转移。因此，它可能为HCC提供更有效的基因治疗。少

The many past approaches used to treat hepatocellular carcinoma (HCC) patients have been less than satisfactory. Accumulating evidence has shown that carcinoma cell invasion and metastasis are inhibited by NK4, an antagonist for hepatocyte growth factor (HGF), and that gene therapy using replication-competent adenovirus is an effective treatment for cancer. HCC, approximately 70% of which can produce u-fetoprotein (AFP), is considered as a candidate disease for the development of a novel treatment using different genes and different vectors. To develop a rational basis for effective anticancer therapy, inhibitory effects of treatment with an AFP-promoter-driven adenovirus deficient of the E1B-55k gene (AdAFP/Rep) and an adenoviral vector expressing NK4 (AdCMV.NK4, provided by Prof. T. Nakamura, Osaka University, and Prof. T. Nukiwa, Tohoku University) to target AFP-producing HCC cells on the tumor growth and invasion were examined in vivo and in vitro. AdAFP/Rep was able to lyse all th … More e AFP-producing HCC cell line (HuH7, HepG2) and not to lyse normal human hepatocytes in primary culture. AdCMV.NK4 played an inhibitory role in the proliferation of the AFP-producing HCC cells, but not normal hepatocytes. Invasion of AFP-producing HCC cells through Matrigel basement membrane components and into collagen gels was inhibited by treatment with AdCMV.NK4. Treatment with AdAFP/Rep or AdCMV.NK4 in nude mice implanted AFP-producing HCC cells resulted in an inhibition in the tumor growth. AdCMV.NK4 treatment decreased mRNA expressions of vascular endothelial growth factor (VEGF) and CD31 as well as matrix metalloproteinase(MMP)-2 and MMP-9 in the HCC cells and implanted subcutaneous tumors. Although AdCMV.NK4 alone did not have a more potent stimulatory effect on apoptosis than that of AdAFP/Rep, the combined treatment with AdAFP/Rep and AdCMV.NK4 induced synergic pro-apoptotic action in the cells and tumors, and inhibited growth and metastasis of invisible HCC much more than each treatment. These findings clearly showed that the combination of AdAFP/Rep and AdCMV.NK4 can inhibit tumor growth, invasion and metastasis of HCC cells in laboratory animals. It may therefore offer a more effective gene therapy for HCC. Less