探讨联合应用凋亡素(apoptin)基因、新城疫病毒(newcastle disease virus, NDV)血凝素神经氨酸酶（hemagglutininneuramidinase, HN）基因及人白介素18（hIL18）基因体外对人结肠癌细胞HCT116的抑制效应。方法：重组质粒pIRApoptinHNIL18通过脂质体介导法体外转染人结肠癌细胞HCT116，通过Western blotting和RTPCR检测外源基因表达；采用AO/EB染色法检测pIRApoptinHNIL18对人结肠癌细胞HCT116抑制作用；利用流式细胞术分析pIRApoptinHNIL18对人结肠癌细胞HCT116线粒体膜电位(mitochondrial transmembrane potential，△Ψ)和活性氧(reactive oxygen species，ROS)水平；采用3,5二羟基甲苯法测定唾液酸含量。结果：pIRApoptinHNIL18转染人结肠癌细胞HCT116后，外源基因能够有效表达；肿瘤细胞生长受到抑制；线粒体△Ψ由(94.41±8.17)%下降到(30.70±8.01)%(P<0.05)；唾液酸含量由(0.33±0.06) mmol/L下降到(0.09±0.03) mmol/L(P<0.01)；ROS水平由(52.48±6.09)%升高到(68.98±7.26)%(P<005)。结论：pIRApoptinHNIL18可通过线粒体途径诱导细胞凋亡，从而抑制人结肠癌细胞HCT116的生长。

To investigate the combined antitumor effects of apoptin gene, hemagglutininneuramidinase gene (HN) of newcastle disease virus (NDV), and human IL18 gene(hIL18) on colon carcinoma cells HCT116in vitro. Methods:The recombinant plasmid pIRApoptinHNIL18 was introduced into human colon carcinoma cells HCT116 by liposomemediated transfection. The expression of the exogenous genes was detected by Western blotting and RTPCR. The antitumor effects of pIRApoptinHNIL18 on HCT116 cells were determined by AO/EB staining. The influence of pIRApoptinHNIL18 on mitochondrial transmembrane potential (△Ψ) and reactive oxygen species (ROS) were detected by flow cytometry (FCM). Sialic acid content was determined by using 3, 5dihydroxytoluene. Results: pIRApoptinHNIL18 transfection resulted in expression of the exogenous genes, repression of HCT116 cells, downregulation of △Ψ \[(94.41±8.17) % vs (30.70±8.01)%,P<005\] and Sialic acid content \[(0.33±0.06) mmol/L vs (0.09±003) mmol/L,P<0.01\], and upregulation of ROS \[(52.48±6.09)% vs (68.98±7.26)% P<0.05\]. Conclusion: pIRApoptinHNIL18 may inhibit the growth of HCT116 cells via mitochondrial pathwayinduced apoptosis.

R719.31

吉林省科技发展计划资助项目 (No.20060566)