提高腺病毒对肿瘤细胞转染效率的药物效果评价

Improvement of the Transfection Efficiency of Recombinant Adenovirus on Cancer Cells Bydodecylmaltoside

为了解决在肿瘤基因治疗中腺病毒载体介导的转染细胞效率低的问题,高效、低毒的转染试剂为其提供了一条新途径.采用正十二烷基-β-D-麦芽糖苷(n-Dodecyl-β-D-Maltoside,DDM)、DC-Chol、SDS联合腺病毒e GFP-Ad转染肝癌细胞Hep G2、食管癌细胞EC109及人正常肝细胞L-02与食管正常细胞Het-1A,48 h后观察绿色荧光蛋白表达情况并计阳性细胞率,评价联合转染试剂处理提高e GFP-Ad转染肿瘤细胞的效率.实验结果表明,DDM(3μg/m L)、DC-Chol(1 000μg/m L)、SDS(0.5μg/m L)联合e GFP-Ad转染肿瘤细胞Hep G2和EC109的效率分别可达85%和78%、80%和76%、45%和41%,与单独e GFP-Ad相比,转染效率分别提高16倍和14.6倍、15倍和14.2倍、8倍和7.2倍;且与正常细胞相比,DDM能显著提高对肿瘤细胞的转染效率,二者差异具有高度统计学意义.作为非脂质体类试剂,DDM能显著提高腺病毒对肿瘤细胞的转染效率,为进一步提高腺病毒介导的肿瘤基因治疗提供有效方法与技术参考.

In order to solve the problem of low adenovirus-transfected efficiency on cancer cells of tumor gene therapy, efficient and low-toxicity transfection reagents have been considered as a promising method. In this research, n-Dodecyl-β-D-Maltoside( DDM), DC-Chol and SDS were affected synergistically with e GFP-Ad to transfect HepG2,EC109,L-02 and Het-1 A cells. The drug effects were evaluated by detection and comparison of the green fluorescence expression after 48 hours of adenovirus transfection. The data showed that the transfection efficiency of DDM, DC-Chol, and SDS was significantly improved in HepG2 and EC109 cancer cells when comparing to L-02 and Het-1 A human normal cells. The incremental transfect efficiencies of DDM( 3 μg/m L),DC-Chol( 1 000 μg/m L) and SDS( 0. 5 μg/m L) on tumor cells HepG2 and EC109,at about 85% and 78%,80% and 76%,45%and 41%,were 16 times and 14. 6 times,15 times and 14. 2 times,8 times and 7. 2 times as great as the non-treated groups. The results indicated that DDM significantly improved the transfection efficiency of adenovirus on tumor cells,which can provide the technical reference for adenovirus-mediated gene therapy.