晚期肝细胞癌（HCC）仍然是一种无法治愈的疾病，总体生存期不到一年。缺氧诱导因子 1α (HIF-1α) 水平升高是导致治疗抵抗力增加和复发的关键分子介质之一，从而导致肿瘤细胞转移。已知多种 microRNA 失调并影响 HCC 中的 HIF-1α 表达。计算机分析表明 hsa-miR-199a-5p 在 HCC 的各个阶段均下调，并且已知可抑制 HIF-1α 表达。基于此分析，我们通过使用编码诱导型半胱天冬酶 9 (iCasp9) 和 miR-199a 的亲肝性腺相关病毒载体开发了一种组合自杀基因疗法。单独过表达 miR-199a-5p 显着降低（与模拟处理细胞相比 ~ 2 倍，p< 0.05）HIF-1α mRNA 水平，同时体外 Huh7 细胞和异种移植模型中癌细胞的细胞毒性增加。体内。为了进一步增强基因治疗的疗效，我们评估了 AAV8-miR-199a 和 AAV6-iCasp9 在 HCC 异种移植模型中的协同治疗效果。我们的数据显示，接受联合自杀基因治疗的小鼠表现出 HIF-1α 表达降低（与模拟治疗相比， ~ 4 倍，p< 0.001），与模拟治疗的动物相比，肿瘤生长显着减少。这些发现强调了 HCC 自杀基因治疗期间下调 HIF-1α 的治疗潜力。© 2024。作者获得 Springer Nature America, Inc. 的独家许可。

Advanced-stage hepatocellular carcinoma (HCC) remains an untreatable disease with an overall survival of less than one year. One of the critical molecular mediators contributing to increased resistance to therapy and relapse, is increased hypoxia-inducible factor 1α (HIF-1α) levels, leading to metastasis of tumor cells. Several microRNAs are known to be dysregulated and impact HIF-1α expression in HCC. An in silico analysis demonstrated that hsa-miR-199a-5p is downregulated at various stages of HCC and is known to repress HIF-1α expression. Based on this analysis, we developed a combinatorial suicide gene therapy by employing hepatotropic Adeno-associated virus-based vectors encoding an inducible caspase 9 (iCasp9) and miR-199a. The overexpression of miR-199a-5p alone significantly decreased ( ~ 2-fold vs. Mock treated cells, p < 0.05) HIF-1α mRNA levels, with a concomitant increase in cancer cell cytotoxicity in Huh7 cells in vitro and in xenograft models in vivo. To further enhance the efficacy of gene therapy, we evaluated the synergistic therapeutic effect of AAV8-miR-199a and AAV6-iCasp9 in a xenograft model of HCC. Our data revealed that mice receiving combination suicide gene therapy exhibited reduced expression of HIF-1α ( ~ 4-fold vs. Mock, p < 0.001), with a significant reduction in tumor growth when compared to mock-treated animals. These findings underscore the therapeutic potential of downregulating HIF-1α during suicide gene therapy for HCC.© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.