~(113)In~m-p-SCN-Bz-DTPA-c-myc反义核酸的合成及其抑制血管平滑肌细胞增殖
Synthesis of ~(113)In~m-p-SCN-Bz-DTPA-c-myc Antisense Oligonucleotides and Investigation of Its Inhibition Effects on the Proliferation of Vascular Smooth Muscle Cells
-
摘要: 为研究113Inm-p-SCN-Bz-DTPA-c-myc反义核酸对血管平滑肌细胞增殖的影响,合成了c-myc反义核酸,与双功能团连接剂p-SCN-Bz-DTPA偶联后又进行了113Inm标记,考察了标记物113Inm-p-SCN-Bz-DTPA-c-myc反义核酸在生理盐水和牛血清的体外稳定性、杂交能力和对平滑肌细胞增殖的影响。测得标记率为45%~55%,纯化后放化纯>90%。标记物在生理盐水和牛血清的体外稳定性均较好,48 h后放化纯>95%。杂交实验中放射性反义链和正义链杂交结合形成大分子,说明偶联和标记没有破坏放射性反义链杂交反应活性。细胞增殖实验显示,113Inm-p-SCN-Bz-DTPA-c-myc反义核酸对平滑肌细胞增殖的抑制率具有剂量依赖性,浓度为10μmol/L(2.5×37 GBq/L)时抑制率最大;113Inm-p-SCN-Bz-DTPA-c-myc反义核酸(放射性反义治疗组)和反义核酸(反义治疗组)1、13InmCl3溶液(放射性治疗组)、不加核酸或113InmCl3溶液(对照组)和c-myc正义核酸(正义治疗组)对平滑肌细胞增殖的抑制率分别为94.92%,69.28%,20.55%,15.24%,0%,放射性反义治疗组和反义治疗组、放射性治疗组、对照组比较有显著性差异(p<0.01),反义治疗组和正义治疗组比较具有显著性差异(p<0.01)。说明113Inm-p-SCN-Bz-DTPA-c-myc反义核酸在体外可有效抑制猪血管平滑肌细胞增殖。Abstract: To investigate the effect of ~(113)In~m-pSCN-Bz-DTPA-c-myc antisense oigonucleotides(ASON) on the proliferation of vascular smooth muscle cells(VSMCs),the c-myc ASON(3'-cholestyl-TAC GGG GAG TTG CAA-C3 NH_2 5',3' end has been sulfurized) is synthesized and conjugated with the chelating agent p-SCN-Bz-DTPA,and then labeled with ~(113)In~m.The labeling yield is 45%~55%,and the radiochemical purity is higher than 90%.The stability of ~(113)In~m-DTPA-c-myc ASON in saline and in bovine serum is good,as indicated by its radiochemical purity still higher than 90% for 24 h.The hybridization ability of the radiolabeled ASON with its sense oigonucleotide(SON) counterpart(de)-monstrates its bioactivity not damaged during the synthesis.Its inhibition of the proliferation of VSMCs is dose dependent.The best inhibiton occurs at a dose of 10 μmol/L(92.5 GBq/L).The(inhibition) rates of ~(113)In~m-DTPA-c-myc ASON(radiolabeled ASON-treated group),ASON(ASON-treated group),~(113)In~mCl_3 solution(radionuclide-treated group),blank(control group),and SON(SON-treated group) are 94.92 %,69.28 %,20.55 %,15.24 % and 0 %,respectively.The inhibtion by radiolabeld ASON treatment is significantly higher(p<0.01) than that of all other treatments.These experimental results show ~(113)In~m-p-SCN-Bz-DTPA-c-myc antisense oigonucleotides can effectively inhibit the proliferation of VSMCs in vitro.
下载: