Zhang Haitao, Qin Ling, Li Chaolu, Jiang Jianyi, Sun Libo, Zhao Xiaofei, Li Ning
Department of General Surgery, Beijing Youan Hospital, Capital Medical University, Beijing 100069, P.R. China.
Department of Biomedical Information Center, Beijing Youan Hospital, Capital Medical University, Beijing 100069, P.R. China.
Exp Ther Med. 2019 Mar;17(3):1649-1655. doi: 10.3892/etm.2019.7147. Epub 2019 Jan 2.
The present study aimed to examine the apoptotic effects of adenovirus (ADV)-mediated herpes simplex virus thymidine kinase (ADV-TK) combined with ganciclovir (GCV) in tissues obtained from patients with hepatocellular carcinoma in order to provide a theoretical basis for the development of this gene therapy program. Apoptosis detection was conducted using the terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling assay and the apoptosis index was compared between the experimental; and control groups. Furthermore, the protein expression levels of caspase-3, B-cell lymphoma-2 (Bcl-2), Bcl-2-assoicated protein X (Bax) and nuclear factor (NF)-κB were examined in pathological specimens using immunohistochemical staining. The Bax/Bcl-2 ratio and the release of cytochrome were examined using western blot analysis. Results indicated that combined ADV-TK and GCV treatment significantly increased the number of apoptotic cells compared with the control group (P<0.05). Immunohistological analysis revealed that ADV-TK and GCV treatment significantly increased the number of caspase-3-positive cells, reduced the Bax/Bcl-2 ratio and NF-κB expression levels and promoted the release of cytochrome compared with the control group (P<0.01). In conclusion, the present results suggest that combined ADV-TK and GCV treatment exerts its effect through the apoptotic signaling pathway.
本研究旨在检测腺病毒（ADV）介导的单纯疱疹病毒胸苷激酶（ADV-TK）联合更昔洛韦（GCV）对肝癌患者组织的凋亡作用，为该基因治疗方案的开发提供理论依据。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法进行凋亡检测，并比较实验组和对照组的凋亡指数。此外，使用免疫组织化学染色法检测病理标本中半胱天冬酶-3、B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关蛋白X（Bax）和核因子（NF）-κB的蛋白表达水平。采用蛋白质印迹分析检测Bax/Bcl-2比值和细胞色素的释放。结果表明，与对照组相比，ADV-TK和GCV联合治疗显著增加了凋亡细胞数量（P<0.05）。免疫组织学分析显示，与对照组相比，ADV-TK和GCV治疗显著增加了半胱天冬酶-3阳性细胞数量，降低了Bax/Bcl-2比值和NF-κB表达水平，并促进了细胞色素的释放（P<0.01）。总之，目前的结果表明，ADV-TK和GCV联合治疗通过凋亡信号通路发挥作用。
