重组的r-Caspsae-3促凋亡基因体内外治疗胰腺癌的试验研究

目的 观察重组活化的Caspsae-3(r-Caspase-3)促凋亡基因对胰腺癌细胞(PC-Ⅱ)及裸鼠皮下种植瘤的治疗作用。方法 采用先前构建的r-Caspase-3促凋亡基因转染胰腺癌细胞，细胞形态学、DNA电泳、流式细胞学观察胰腺癌细胞凋亡状况；用脂质体-pcDNA3．1( )／r-Caspase-3复合物，治疗人胰腺癌细胞裸鼠种植瘤，观察肿瘤生长速度并计算肿瘤体积大小。结果 DNA电泳、流式细胞学可分别观测到特征性的DNA Ladder和凋亡峰，皮下种植肿瘤治疗后生长速度开始减缓，组织切片HE染色可见散在凋亡细胞；对照组肿瘤生长速度明显快于治疗组。结论 r-Caspase-3具有自催化凋亡诱导活性，以r-Caspase-3为目的基因的基因治疗可能为治疗胰腺癌以及其它恶性肿瘤开辟新的途径。

Therapeutic effects of recombinant caspase-3 gene on human pancreatic carcinoma in vivo and in vitro

Objective To study the inhibitory effects of the recombinant caspases-3 gene (r-caspases-3) eukaryotic expression plasmid on growth of pancreatic carcinoma in vivo and in vitro. Methods Human pancreatic carcinoma cells were infected with r-caspases-3 and the cell apoptosis observed by cell morphology, DNA electrophoresis and flow cytometry, respectively. The model of pancreatic carcinoma subcutaneously transplanted in nude mice was established and subsequently divided into 2 groups: pcDNA3.1(+)/r-caspases-3 group and PBS group. Different treatments were given. The volume of subcutaneous tumor was measured and the morphological changes of tumor observed under microscope. Results r-caspases-3 mRNA could express in infected cells and induced occurrence of apoptosis. The volume was 0.78 cm3 in the pcDNA3.1(+)/r-caspases-3 group and 1.21 cm 3 in PBS group. In the former group, a lot of apoptotic cells were seen in tissue section by HE staining. Conclusions r-caspases-3 gene has autocatalytical activity of apoptosis and may provide a new way for treatment of pancreatic carcinoma.