论文部分内容阅读
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) can enhance the in vivo therapeutic effect of herpes simplex virus thymidine kinase (HSV-TK) suicide gene on hepatocellular carcinoma (HCC). Methods Human hepatocellular carcinoma SMMC-7721 cells were used to establish subcutaneous xenograft model in nude mice. The antitumor effects of different doses of ATRA were detected firstly. Combined with ATRA gene therapy, the experiment was divided into four groups: phosphate buffer group, ATRA group and HSV-TK group , The combined treatment group, the tumor volume was measured in each group, the tumor inhibition rate was calculated, the expression of the target gene plasmid was detected by Western blot, the pathological observation was performed by HE staining and the expression of connexin Cx32 was detected by immunohistochemistry. Results ATRA had no anti-hepatocarcinoma effect at 1 mg / kg, but had a significant anti-hepatoma effect at ≥4 mg / kg. The inhibition rate of ATRA in phosphate buffered saline group, ATRA group, HSV-TK group and combined treatment group was 0 , (7.46 ± 7.25)%, (37.65 ± 3.87)% and (59.04 ± 3.08)%, respectively. There was significant difference between the combination therapy group and phosphate buffer group, ATRA group and HSV-TK group (P < 0.05). The results of HE showed that after treatment with ATRA, the number of tumor necrosis cells was significantly higher than that of the control group. The results of immunohistochemistry showed that the expression of Cx32 protein was increased in ATRA-treated tissues, and the expression of Cx32 was mainly in the membrane. CONCLUSION: ATRA has an anti-hepatocarcinoma effect. ATRA can enhance the effect of HSV-TK suicide gene encapsulated by ultrasound microbubbles on hepatocellular carcinoma. The possible mechanism is that ATRA can promote the normal expression of Cx32 protein.