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目的观察靶向性重组腺病毒Ad-EAFP-PALB/r-Caspase-3对原发性肝癌HepG2细胞及裸鼠皮下种植瘤的治疗作用。方法采用先前构建的靶向性重组腺病毒Ad-EAFP-PALB/r-Caspase-3感染肝癌HepG2细胞、乳腺癌MDA-MB-231细胞以及正常肝L-02细胞,观察细胞形态学改变,并通过流式细胞术检测细胞凋亡指数;采用裸鼠皮下种植瘤模型、重组腺病毒瘤内给药治疗,观察肿瘤生长情况及病理学改变。结果 流式细胞学可见HepG2细胞凋亡和明显的凋亡峰,MDA-MB-231和L-02细胞凋亡指数分别为0和7.3%,明显低于HepG2细胞(48.2%),P<0.01。种植肿瘤细胞2周后HepG2细胞组、MDA-MB-231细胞组和对照组肿瘤体积大小分别为(0.26±0.31)cm3、(0.25±0.15)cm3和(0.26±0.28)cm3,各组间差异无统计学意义(P>0.05);种植肿瘤细胞4周后HepG2细胞组、MDA-MB-231细胞组和对照组肿瘤体积大小分别为(0.53±0.12)cm3、(0.49±0.22)cm3和(0.54±0.13)cm3,各组间差异无统计学意义(P>0.05);HepG2细胞组肿瘤经重组腺病毒Ad-EAFP-PALB/r-Caspase-3治疗后8周时肿瘤体积为(0.65±0.13)cm3,明显小于MDA-MB-231细胞组〔(1.27±0.32)cm3〕和对照组〔(1.43±1.09)cm3〕,P<0.01,而后两组之间比较差异无统计学意义(P>0.05)。治疗后HepG2细胞组肿瘤细胞出现坏死区,伴淋巴浸润细胞和凋亡细胞;对照组肿瘤细胞区域内未见明显坏死区;而MDA-MB-231细胞组肿瘤细胞呈实性排列,呈小腺腔样结构,异型性明显,瘤巨细胞及多核瘤细胞可见,治疗前、后未见明显变化。结论 靶向性重组腺病毒Ad-EAFP-PALB/r-Caspase-3具有有效靶向性凋亡诱导作用,体内、外治疗原发性肝癌有效,可能为原发性肝癌的靶向性基因治疗提供新途径。
Objective To observe the therapeutic effect of targeted recombinant adenovirus Ad-EAFP-PALB / r-Caspase-3 on primary hepatocellular carcinoma HepG2 cells and subcutaneous xenografts in nude mice. Methods HepG2 cells, breast cancer MDA-MB-231 cells and normal liver L-02 cells were infected with the targeted recombinant adenovirus Ad-EAFP-PALB / r-Caspase-3 and the morphological changes were observed The apoptosis index was detected by flow cytometry. The subcutaneous tumor model was established in nude mice and the recombinant adenovirus was administered intraperitoneally to observe the tumor growth and pathological changes. Results Apoptosis and apoptotic peak of HepG2 cells were observed by flow cytometry. The apoptotic index of MDA-MB-231 and L-02 cells were 0 and 7.3%, which was significantly lower than that of HepG2 cells (48.2%, P <0.01) . The tumor volume of HepG2 cell group, MDA-MB-231 cell group and control group were (0.26 ± 0.31) cm3, (0.25 ± 0.15) cm3 and (0.26 ± 0.28) cm3 respectively after planting tumor cells for 2 weeks. The tumor volume of HepG2 cells, MDA-MB-231 cells group and control group were (0.53 ± 0.12) cm3, (0.49 ± 0.22) cm3 and 0.54 ± 0.13) cm3, the difference was not statistically significant (P> 0.05). The tumor volume in HepG2 cells treated with recombinant adenovirus Ad-EAFP-PALB / r-Caspase- 0.13) cm3, which was significantly lower than that of MDA-MB-231 cells [(1.27 ± 0.32) cm3〕 and control group (1.43 ± 1.09 cm3〕, P <0.01, but there was no significant difference between the two groups > 0.05). After treatment, the tumor cells in HepG2 cell group showed necrotic area, with lymph infiltrating cells and apoptotic cells; there was no obvious necrotic area in the tumor cells in the control group; however, the tumor cells in the MDA-MB-231 cells group were arranged in solid, Cavity-like structure, atypia obvious tumor giant cells and polycystic tumor cells can be seen before treatment, no significant change. Conclusion Targeted recombinant adenovirus Ad-EAFP-PALB / r-Caspase-3 can effectively induce apoptosis in vitro and in vivo and in vitro for the treatment of primary liver cancer, which may be targeted gene therapy for primary liver cancer Provide new ways.