Beijing Institute of Hepatology and Beijing YouAn Hospital, Capital Medical University, Beijing 100069, P.R. China.
Int J Oncol. 2017 Oct;51(4):1291-1299. doi: 10.3892/ijo.2017.4105. Epub 2017 Aug 29.
The present study was designed to investigate the synergistic inhibitory effects on hepatocellular carcinoma with recombinant human adenovirus Aspp2 (Aspp2-ad) and oxaliplatin via p53-independent pathway in vitro and in vivo. After being treated with Aspp2-ad and/or oxaliplatin for 24-48 h, HepG2P53-/- and Hep3B cells showed a significant growth inhibition compared with vehicle control. Combination group showed a synergetic effect, the inhibitory rates were all above 80% at 48 h point in HepG2P53-/- and Hep3B cells. The apoptotic cell numbers of Aspp2-ad and/or oxaliplatin treatment groups were increased remarkably, especially for the combined therapy group in the liver cancer cells. The Hep3B xenograft experiment also showed similar inhibition of Aspp2-ad and/or oxaliplatin to the in vitro experiment. H&E results showed that combination group had the least mitotic indexes and the most necrosis. The immunohistochemistry results showed that PCNA, CD31 expression decreased greatly in treatment groups. These results suggested that Aspp2-ad might inhibit proliferation and vascular growth of hepatocarcinoma. Aspp2 induced apoptosis protein expression in Aspp2-ad and combination groups, the Aspp2, Bax and activation of caspase-3 expression increased greatly both in vitro and in vivo. But interestingly, the autophagy proteins showed different responses not only in HepG2P53-/- and Hep3B cells but also in vitro and in vivo. We found that Aspp2-ad downregulated the p-ERK, p-STAT3 expression, the synergistic effects were observed in combination group, while there was not response of mTOR to Aspp2-ad. In conclusion, Aspp2-ad, in P53-independent manner, regulated ERK and STAT3 signal moleculars to inhibit hepatocarcinoma in coordination with oxaliplatin by influencing the protein expression of proliferation, apoptosis, autophagy and vascular growth. Aspp2-ad has the potential to be developed in gene therapy for HCC, especially for P53 deletion or mutation in HCC.
本研究旨在探讨重组人腺病毒 Aspp2（Aspp2-ad）与奥沙利铂通过 p53 非依赖性途径在体外和体内对肝癌协同抑制作用。经 Aspp2-ad 和/或奥沙利铂处理 24-48 小时后，HepG2P53-/-和 Hep3B 细胞的生长抑制率与载体对照组相比均有显著差异。联合组表现出协同作用，在 HepG2P53-/-和 Hep3B 细胞中，48 小时点的抑制率均超过 80%。Aspp2-ad 和/或奥沙利铂处理组的凋亡细胞数量明显增加，尤其是联合治疗组。肝癌细胞中。Hep3B 异种移植实验也显示出与体外实验相似的 Aspp2-ad 和/或奥沙利铂抑制作用。H&E 结果显示，联合组的有丝分裂指数最低，坏死最多。免疫组化结果显示，PCNA、CD31 表达在治疗组中显著降低。这些结果表明，Aspp2-ad 可能抑制肝癌的增殖和血管生长。Aspp2 在 Aspp2-ad 和联合组中诱导凋亡蛋白表达，Aspp2、Bax 和 caspase-3 激活表达在体外和体内均显著增加。但有趣的是，自噬蛋白的反应不仅在 HepG2P53-/-和 Hep3B 细胞中，而且在体外和体内均不同。我们发现，Aspp2-ad 下调了 p-ERK、p-STAT3 的表达，在联合组中观察到协同作用，而 mTOR 对 Aspp2-ad 没有反应。总之，Aspp2-ad 通过影响增殖、凋亡、自噬和血管生长相关蛋白的表达，以 P53 非依赖性方式调节 ERK 和 STAT3 信号分子，与奥沙利铂协同抑制肝癌。Aspp2-ad 有可能开发为 HCC 的基因治疗，特别是在 HCC 中 P53 缺失或突变的情况下。
