摘要
目的探讨全反式维甲酸(all-trans retinoic acid,ATRA)能否提高微泡包裹的单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,HSV-TK)自杀基因对肝癌的体内治疗作用。方法用人肝癌SMMC-7721细胞建立皮下裸鼠移植瘤模型,先检测不同剂量ATRA的抗肿瘤效应;再用ATRA联合基因治疗,实验分4组:磷酸盐缓冲液组、ATRA组、HSV-TK组、联合治疗组,各组测量肿瘤体积,计算抑瘤率,Western blot检测输注的目的基因质粒的表达,HE染色行病理学观察,免疫组化法检测连接蛋白Cx32的表达。结果 ATRA在1 mg/kg下无抗肝癌作用,但≥4 mg/kg即具有明显抗肝癌效应;抑瘤率在磷酸盐缓冲液组、ATRA组、HSV-TK组、联合治疗组分别为0、(7.46±7.25)%、(37.65±3.87)%、(59.04±3.08)%,联合治疗组分别与磷酸盐缓冲液组、ATRA组、HSV-TK组比较,差异有统计学意义(P<0.05)。HE结果显示:采用ATRA联合基因治疗后,肿瘤坏死细胞明显较对照组多,免疫组化结果显示:ATRA处理过的组织中,Cx32蛋白表达增加,且以胞膜表达为主。结论 ATRA具有抗肝癌作用;ATRA能增强超声微泡包裹的HSV-TK自杀基因杀伤肝癌的效果,其可能机制与ATRA促进了Cx32蛋白的正常表达有关。
Objective To investigate whether all-trans retinoic acid（ATRA） could enhance the killing effect of ultrasound microbubble carrying herpes simplex virus thymidine kinase（HSV-TK） on hepatocellular carcinoma in vivo.Methods Nude mice were inoculated subcutaneously with SMMC-7721 tumor cells.The anti-tumor effect of ATRA at different doses was tested by measuring the tumor size to obtain a dose without anti-tumor effect.SMMC-7721 tumor cells transplanted nude mice were randomized into 4 groups,PBS treatment group,ATRA treatment group,HSV-TK treatment group,combined treatment group.The corres-ponding treatments were given by tail injection at 0.2 ml/time,once per 3 d,for 15 d.The later 2 groups received an ultrasonic irradiation at 3 000 Hz,2 W/cm2 for 5 min.ATRA was given by intraperitoneal injection at a dose of 1 mg/（kg·d） for 15 d.Plasmid gene was test by Western blotting.The tumors were removed for histopathological analysis.Protein Cx32 was detected by immunohistochemistry.Results ATRA showed no anti-tumor effect at a dose of 1 mg/kg,but it showed anti-tumor effect when the dose beyond 4 mg/kg.The inhibitory rate of tumor growth was 0 in PBS-treat group,（7.46±7.25）% in ATRA treatment group,（37.65±3.87）% in HSV-TK treatment group,and（59.04±3.08）% in combined treatment group（F=116.709,P0.05;For combined treat group vs PBS treatment group,ATRA treatment group,HSV-TK treatment group,t=59.04,51.55,and 21.39,respectively,P0.05）.HE staining showed that extensive necrosis was observed in combined treatment group when compared with the other groups.The expression of protein Cx32 were increased in the groups with ATRA treatment and mainly located in the cell membrane.Conclusion ATRA enhances the killing effect of ultrasound microbubble carrying HSV-TK on hepatocellular carcinoma in vivo,which may be associated with promoting the expression of Cx32 by ATRA.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第20期2112-2115,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30872977)~~
