^(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

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。

To investigate the effect of ^113In^m-p-SCN-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 NH2 5＇, 3＇ end has been sulfurized） is synthesized and conjugated with the chelating agent p-SCN-Bz-DTPA, and then labeled with rainm. The labeling yield is 45%～55%, and the radiochemical purity is higher than 90%. The stability of ^113In^m-DTPA-c-myc ASON in saline and in bovine serum is good, as indicated by its radiochemical purity still higher than 90% for 24h. The hybridization ability of the radiolabeled ASON with its sense oigonucleotide （SON） counterpart demonstrates 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 ^113In^m-DTPA-c-myc ASON （radiolabeled ASON-treated group）, ASON （ASON-treated group）, ^113In^mCl3 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 ^113In^m-p-SCN-Bz-DTPA-c-myc antisense oigonucleotides can effectively inhibit the proliferation of VSMCs in vitro.