摘要
目的 评价99Tcm 标记血管活性肠肽 (VIP)受体介导的C mycmRNA反义寡核苷酸复合物 (99Tcm VIP ASON)对结肠腺癌HT2 9细胞的反义抑制作用。方法 用99Tcm 标记长度为 15个碱基互补于C mycmRNA的反义寡核苷酸 (ASON) ,在一定条件下与VIP形成99Tcm VIP ASON复合物 ,测定HT2 9细胞对99Tcm VIP ASON的摄取率以及99Tcm VIP ASON对HT2 9细胞生长和C myc癌蛋白质表达的影响。结果 摄取研究显示 ,各时间点HT2 9细胞对99Tcm VIP ASON的摄取均高于99Tcm ASON(P <0 .0 5 ) ,摄取率在 12 0min达最高 ,为 2 7.39%。噻唑蓝 (MTT)比色试验中 ,99Tcm VIP ASON组在各剂量下吸光度值均明显低于99Tcm ASON和99Tcm 正义寡核苷酸 (SON)、99Tcm 无义寡核苷酸 (MON)、VIP 多聚赖氨酸结合物 (VIP PL)及空白对照组 ,流式细胞仪测定99Tcm VIP ASON组C myc癌蛋白质的荧光强度为 0 6 33± 0 0 38,显著低于其他各组 (P <0 .0 1)。结论 VIP受体介导途径能显著增加99Tcm ASON的转染效率 ,明显抑制HT2 9细胞的生长和C myc癌蛋白质的表达 ,增强反义抑制作用效果。
Objective To explore the antisense inhibition effect of vasoactive intestinal peptide (VIP) receptor mediated radiolabeled C myc mRNA antisense oligonuclide complexes ( 99 Tc m VIP ASON) on the human colon adenocarcinoma cell line HT29. Methods A 15 base single stranded antisense oligonuclide (ASON) complementary to C myc mRNA was labeled with 99 Tc m. 99 Tc m ASON was complexed with VIP under certain condition. The HT29 cells were incubated with the complex and the uptake rate of 99 Tc m VIP ASON was assayed. The effect of 99 Tc m VIP ASON on cell growth and C myc cancer protein expression was studied by VIP receptor mediated endocytosis. Results The uptake rate of HT29 cell exposed to 99 Tc m VIP ASON complex was highly increased by VIP receptor mediated endocytosis, the highest uptake rate was 27.39% at 2 h and significantly higher than that exposed to 99 Tc m ASON ( P <0.05).The absorbance tested by methyl thiazol tetrazolium (MTT) assay in the 99 Tc m VIP ASON group was significantly lower than in other study groups ( P <0.05). The expression of C myc cancer protein detected by flow cytometry was 0.633±0.038 in the 99 Tc m VIP ASON group and was very significantly lower than that in other groups ( P <0.01). Conclusion VIP receptor mediated endocytosis pathway could highly increase the access of 99 Tc m ASON into HT29 cell and the 99 Tc m ASON entering into cells could effectively inhibit cell proliferation and C myc cancer protein expression.
出处
《中华核医学杂志》
CAS
CSCD
北大核心
2003年第3期148-151,共4页
Chinese Journal of Nuclear Medicine
基金
国家自然科学基金资助项目 (39870 2 0 0 )
