自稳定反义IGF1R基因片段对膀胱癌细胞的影响

Effects of self-stablized antisense oligodeoxynucleotide targeted against IGF1R gene on T24 bladder cancer cells

目的 探讨针对胰岛素样生长因子 1受体 (IGF1R )基因的自稳定反义脱氧寡核苷酸(ODN)对膀胱癌细胞的影响。 方法 采用RT -PCR、Westernblot、MTT试验、流式细胞仪对经反义ODN作用后的T2 4膀胱癌细胞进行动态分析。 结果 自稳定反义ODN在血清培养基中 2 4h内均保持稳定状态 ,而硫代反义ODN 4h后基本降解。自稳定反义ODN可有效降低T2 4膀胱癌细胞IGF1R基因及蛋白表达 ,增强膀胱癌细胞对丝裂霉素的敏感性及凋亡易感性。

Objective To evaluate the effects of self stablized antisense oligodeoxynucleotide (ODN) targeted against insulin like growth factor 1 receptor(IGF1R) gene on T24 bladder cancer cells. Methods Semiquantitative RT PCR, Western blot , MTT determination, flow cytometry were used to study the effects of self stablized antisense ODN on IGF1R gene expression, protein synthesis, drug sensitivity and apoptosis of T24 cells. Results The self stablized antisense ODN was stable in culture medium for 24 hours, whereas phosphorothiate ODN were almost degradated after 4 hours in the same condition. After treated with self stablized antisense ODN specific for IGF1R gene, the intrinsic mRNA expression of IGF1R gene were reduced approximately by 84.3%(phosphorothiate ODN: 74.0%), the protein synthesis of IGF1R were downregulated by 77.0% (phosphorothiate ODN: 61.3%), which caused targeted cells to be more sensitive to mitomycin induced apoptosis (P<0.05). Conclusions These data showed antisense ODN with ability of anti degradation could effectively reduce IGF1R gene expressions and protein synthesis of T24 cells and could significantly enhance apoptotic sensitivity of T24 cells to bladder instilation drug mito mycin.These suggested that IGF1R antisense ODN may serve as a potential therapeutic approach to bladder cancer.