超微载体介导反义端粒酶RNA抑制胶质瘤细胞生长的研究

Nanoparticles mediated antisense telomerase RNA oligonucleotide inhibited the growth of gliomas cells

目的 研究用体内可降解的聚乳酸 (PLA)和o 梭甲基壳聚糖 (CMC)制成的超微载体介导端粒酶RNA(hTR)反义寡核苷酸在体外对TJ90 5人脑胶质瘤细胞的作用。方法 用PLA和CMC制成超微载体 ,并用其介导hTR反义寡核苷酸在体外转染TJ90 5细胞 ,通过噻唑兰比色法(MTT)、改良端粒重复序列扩增法 (TRAP)、逆转录 聚合酶链反应 (RT PCR)对细胞转染情况作检测。结果 超微载体在体外能有效转染hTR反义寡核苷酸 ,48h后细胞存活率为 46.84% ,hTR、端粒酶催化亚基mRNA和端粒酶的活性水平分别为 0 .3 16、0 .0 2 4、5 1.40 0 ,明显受到抑制。结论 hTR可作为胶质瘤基因治疗靶点 ,超微载体能有效转染基因药物 ,可替代病毒载体。

Objective To study the inhibitory effects of antisense human telomerase RNA (hTR) to TJ905 cells (one kind of human glioma cells) mediated by nanoparticles.Methods The nanoparticles were constructed with polylactic acid (PLA) and o-carboxymethylated chitosan (CMC).Fluorescent labeled antisense hTR oligonucleotide was transfected into TJ905 cells through nanoparticles in vitro.The results were identified by MTT assay,TRAP method and RT-PCR method.Results Nanoparticles could transfected antisense hTR oligonucleotide into TJ905 cells effectively.After transfected antisense hTR ODN,proliferation activity of TJ905 cells was inhibited significantly.At 48 h,the survival rate was 46.84%,the telomerase activity was 51.4,and the expression of hTR and hTERT was 0.316 and 0.024 respectively.All were inhibited significantly.Conclusion hTR maybe act as a novel target of gene therapy for malignant gliomas.Nanoparticles could effectively transfect the gene delivery system and substitute for viral carriers.