肿瘤溶解病毒能够选择性地溶解肝脏中的肿瘤细胞，而不会杀死正常的肝细胞，并同时激活免疫反应。肿瘤溶解病毒疗法有望彻底改变肝癌治疗的模式，包括肝细胞癌（HCC）这种最常见和最致命的恶性肿瘤。本研究利用反向遗传学技术，将PR8病毒的NA片段与源自人类端粒酶逆转录酶的GV1001肽进行负载。在体外评估了重组溶瘤病毒的治疗效果之后，又对HCC小鼠进行了体内研究。通过测序验证了重组病毒基因序列，通过血凝试验检测了病毒毒力，基于TCID50确定了病毒毒力。通过电子显微镜观察了病毒的形态结构，并通过细胞免疫荧光法对GV1001肽进行了定位。体外实验表明，重组溶瘤病毒对培养的HCC细胞和正常肝细胞具有选择性细胞毒性，只杀死了肿瘤细胞而未对正常细胞产生显著影响。与体外实验结果一致，重组溶瘤流感病毒在小鼠体内显著抑制了肝肿瘤的生长，同时诱导了抗肿瘤免疫反应，包括CD4+和CD8+T淋巴细胞数量的增加，并改善了小鼠的生存率。我们的结果表明，携带GV1001的溶瘤流感病毒是治疗HCC患者的一种有前景的免疫疗法。

Oncolytic viruses are able to lyse tumor cells selectively in the liver without killing normal hepatocytes, in addition to activating the immune response. Oncolytic virus therapy is expected to revolutionize the treatment of liver cancer, including hepatocellular carcinoma (HCC), one of the most frequent and fatal malignancies. In this study, reverse genetics techniques were exploited to load NA fragments of the PR8 virus with GV1001 peptides derived from human telomerase reverse transcriptase. An in vitro assessment of the therapeutic effect of the recombinant oncolytic virus was followed by an in vivo study in mice with HCC. The recombinant virus was verified by sequencing of the recombinant viral gene sequence, and viral virulence was detected by hemagglutination assays and based on the TCID50. The morphological structure of the virus was observed by electron microscopy, and GV1001 peptide was localized by cellular immunofluorescence. The selective cytotoxicity of the recombinant oncolytic virus in vitro was demonstrated in cultured HCC cells and normal hepatocytes, as only the tumor cells were killed; the normal cells were not significantly altered. Consistent with the in vitro results, the recombinant oncolytic influenza virus significantly inhibited liver tumor growth in mice in vivo, in addition to inducing an anti-tumor immune response, including an increase in the number of CD4+ and CD8+ T lymphocytes and, in turn, improving survival. Our results suggest that oncolytic influenza virus carrying GV1001 is a promising immunotherapy in patients with HCC.