反义PCNA基因片断抑制胶质瘤U251细胞增殖

目的 研究反义PCNA基因片断对胶质瘤细胞株U251的细胞增殖的抑制作用。方法 用脂质体介导寡核苷酸转染培养的U251胶质瘤细胞，以MTT法检测细胞增殖活性，RT-PCR、原位杂交和免疫组织化学方法分别检测相关基因和蛋白的表达，通过流式细胞仪检测细胞周期和TUNEL法检测细胞凋亡。结果 不同浓度的反义寡核苷酸对肿瘤细胞的增殖活性均有抑制作用，且有一定的剂量依赖性。PCNA反义寡核苷酸能够诱导胶质瘤细胞凋亡，且与反义寡核苷酸浓度和作用时间有关。结论 反义PCNA寡核苷酸片断能有效地抑制胶质瘤细胞的体外增殖，且能够诱导胶质瘤细胞的凋亡。

Study of Inhibitory Effect of PCNA Antisense Oligonucleotides on the Proliferation of Glioma U251 Cells

Objective To study the inhibitory effect of proliferating cell nuclear antigen(PCNA) antisene oligonucleotides on the proliferation of glioma cell line U251. Methods To evaluate the effect of PCNA antisense oligonucleotides on tumor cells, ODN was transfected into tumor cell mediated by lipofectin. Then MTT assay was utilized to analyses the effect on the proliferation activity of tumor cells and in situ hybridization, RT PCR and immunohistochemistry used to study expression difference of mRNA and protein after transfection. Cell cycle and cellular apoptosis were also analyzed. Results Tumor cells tended to be inhibited in proliferation and the inhibitory effect seemed to be dose dependent. Both mRNA and protein expression reduced dramatically after transfection. Conclusion PCNA antisene oligonucleotides can greatly reduce the mRNA and protein expression of PCNA, and can inhibit the tumor proliferation, induce apoptosis of glioma tumor cell.