Connexin32‑mediated antitumor effects of suicide gene therapy against hepatocellular carcinoma: In vitro and in vivo anticancer activity.
Connexin32‑mediated antitumor effects of suicide gene therapy against hepatocellular carcinoma: In vitro and in vivo anticancer activity.
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DOI：
10.3892/mmr.2016.4895
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发表时间：
2016-04
影响因子：
3.4
通讯作者：
Lun Wu;Wen-bo Zhou;Feng Shen;Wei Liu;Hong-Wei Wu;Shi-ji Zhou;Sheng-wei Li
中科院分区：
文献类型：
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作者：
Lun Wu;Wen-bo Zhou;Feng Shen;Wei Liu;Hong-Wei Wu;Shi-ji Zhou;Sheng-wei Li
正常肝细胞表达连接蛋白32（Cx 32），其在细胞-细胞接触区域形成间隙连接。本研究的目的是探讨Cx 32是否介导携带单纯疱疹病毒胸苷激酶（HSV-TK）自杀基因的超声微泡在体外和体内对肝癌细胞的细胞死亡诱导作用。采用不同浓度的反式维甲酸（trans-retinoic acid，ATRA）处理HepG 2细胞，观察ATRA的内在抗肿瘤作用。对ATRA通过Cx 32介导的抗肿瘤作用进行了详细的体外和体内研究，然后研究了该化合物可能的潜在作用机制。采用逆转录-定量聚合酶链反应（RT-PCR）方法检测超声波转染后HepG 2细胞中HSV-TK基因的表达。使用MTT测定评估对细胞死亡的影响。通过免疫组织化学分析和Western印迹分析定量ATRA处理或未处理组织中Cx 32的蛋白表达水平。利用超声波法成功地将HSV-TK基因转染到HepG 2细胞中，并获得稳定表达。与其他组相比，ATRA处理的HSV-TK基因组的凋亡细胞数增加（P<0.05），肿瘤抑制作用增强（P<0.05）。ATRA组Cx 32蛋白表达明显高于对照组（P<0.01）。本研究表明ATRA可提高Cx 32蛋白表达，增强HSV-TK/GCV自杀基因治疗系统的旁观者效应，这可能为肝癌的治疗提供一种潜在的策略。
Normal hepatocytes express connexin32 (Cx32), which forms gap junctions at cell‑cell contact areas. The aim of the present study was to investigate whether Cx32 mediates the cell death‑inducing effects of ultrasound microbubbles carrying the herpes simplex virus thymidine kinase (HSV‑TK) suicide gene against hepatocellular carcinoma cells in vitro and in vivo. HepG2 cells were exposed to different concentrations of trans‑retinoic acid (ATRA) in culture, to evaluate the intrinsic antitumor effect of ATRA. Detailed in‑vitro and in‑vivo investigations on the antitumor effects of ATRA via Cx32 mediation were performed, and the possible underlying mechanisms of action of the compound were then examined. The gene expression of HSV‑TK transfected by ultrasound wave irradiation in the HepG2 cells was quantified using reverse transcription‑quantitative polymerase chain reaction analysis. The effects on cell death were assessed using an MTT assay. The protein expression levels of Cx32 in ATRA‑untreated or ATRA‑treated tissues were quantified by immunohistochemical analysis and Western blot assays. The HSV‑TK gene was successfully transfected into the HepG2 cell using ultrasound wave irradiation, and was stably expressed. Compared with the other groups, the HSV‑TK gene group treated with ATRA exhibited an increased number of apoptotic cells (P<0.05) and improved tumor suppression (P<0.05). ATRA significantly increased the expression of Cx32 in the hepatoma tissues (P<0.01). The present study demonstrated that ATRA elevated the protein expression of Cx32 and enhanced the bystander effect of the HSV‑TK/GCV suicide gene therapy system, which may provide a potential strategy for hepatocellular carcinoma treatment.