p53基因治疗联合TACE和射频消融在兔VX2肝癌模型中的疗效研究

Experimental study on the curative effects of p53 gene therapy combined with TACE and radiofrequency ablation on VX2 hepatocellular carcinoma in rabbits

目的探讨p53基因治疗联合经导管动脉化疗栓塞术(TACE)和射频消融对兔VX2肝癌的治疗效果以及对p53蛋白表达的影响。方法24只VX2肝癌模型兔随机分为A、B、C组,每组8只。A组接受瘤内注射rAd-p53,经肝动脉灌注超液态碘油和射频消融治疗,B组接受经肝动脉灌注rAd-p53和超液态碘油及射频消融治疗,C组接受TACE超液态碘油和射频消融治疗。治疗后第3天检测p53蛋白表达量,治疗后1、2、3、4周CT测量肿瘤体积,比较3组治疗前,治疗后1 d、3 d、1周和2周的丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平。最后处死实验兔,观察肿瘤大小及坏死情况并对标本行HE染色。结果治疗4周后组织形态学显示C组标本毁损中心区较A组和B组略小,HE染色显示大片凝固性坏死,凝固性坏死区周围有包膜形成。治疗后1~4周,CT显示A组和B组肿瘤体积呈持续缩小趋势,C组肿瘤体积在治疗后的前2周先缩小,治疗后3周恢复增长。治疗后1周3组肿瘤体积差异无统计学意义(P>0.05),治疗后2、3、4周A、B组肿瘤体积均较C组小,差异有统计学意义(P<0.05)。治疗后1 d 3组ALT和AST水平均升高达到峰值,较治疗前显著升高(P<0.05);治疗后3 d、治疗后1、2周3组的ALT和AST在治疗后1 d的基础上持续下降,其中治疗后3 d、1、2周时,A、B组的ALT和AST水平显著低于C组(P<0.05),治疗后2周时A、B组ALT和AST基本恢复至术前水平,差异有统计学意义(P<0.05)。与C组比较,治疗后3 d A组和B组p53表达更低,差异有统计学意义(P<0.05)。结论p53基因治疗联合TACE和射频消融可抑制肿瘤生长,是兔VX2肝癌的有效治疗方法。

Objective To investigate the therapeutic effects of p53 gene therapy combined with transcatheter artery chemoembolization(TACE)and radiofrequency ablation in treatment of VX2 hepatocellular carcinoma in rabbits,and to observe its effects on p53 protein expression.Methods A total of 24 rabbits with VX2 hepatocellular carcinoma were randomly divided into three groups.The rats in group A received intratumoral injection of rAd-p53,transhepatic arterial infusion of superfluid iodine oil and radiofrequency ablation,and those in group B received transhepatic arterial infusion of rAd-p53 and superfluid iodine oil combined with radiofrequency ablation,and those in group C received TACE superfluid iodine oil and radiofrequency ablation.The expression levels of p53 protein at 3d after treatment were detected,and the tumor volume at 1d,3d,1w and 4w after treatment was measured.Moreover the levels of ALT and AST before treatment,at 1d,3d,1w and 2w after treatment were observed and compared among groups.Finally,the experimental rabbits were executed to observe the tumor size and necrosis status,and the specimens were stained with HE.Results After 4-week treatment,the histomorphology showed that the central area of destruction in group C was slightly smaller than that in groups A and B.HE staining showed a large area of coagulative necrosis with envelope formation around the coagulative necrosis area.CT showed that the tumor volume at 1w~4w after treatment in groups A and B had a continuous reduction trend,while the tumor volume in group C was firstly decreased in the first 2 weeks after treatment,then resumed growth at 3w after treatment.There was no significant difference in tumor volume at 1w after treatment among the three groups(P>0.05),which at 2w,3w and 4w after treatment in groups A and group B was significantly smaller than that in group C(P<0.05).The ALT and AST levels at 1d after treatment in the three groups were significantly increased,however,which were significantly decreased at 3d,1w,2w after treatment,moreover,which in group A and group B were significantly lower than those in group C.In addition the ALT and AST levels in groups A and B at 3d,1w,2w after treatment were basically returned to the levels before operation(P<0.05).Conclusion The p53 gene therapy combined with TACE and radiofrequency ablation can inhibit the tumor growth,which is an effective treatment way for VX2 hepatocellular carcinoma in rabbits.