^(188)Re标记反基因肽核酸的方法学研究

Investigations of Labeling Antigene Peptide Nucleic Acid with ^(188)Re

目的研究188Re标记反基因肽核酸(AGPNA)的方法及标记物与胰腺癌Patu8988细胞结合内化的特性。方法用188Re直接标记经修饰的AGPNA,改变标记条件摸索标记方法,在不同时间点(15min^6h)测定标记率;测定标记物加入人血清和生理盐水后不同时间点(15min^24h)的放化纯度;进行胰腺癌Patu8988细胞摄取188Re-AGPNA的内化实验。结果当标记条件为100μlSnCl2·2H2O(20mg/ml)和20μlAGPNA(2mg/ml)时,188Re-AGPNA的标记率最高可达(89.99±0.15)%,放射性胶体含量为(9.40±0.55)%。标记物加入血清24h后放化纯度为(89.14±0.63)%。188Re-AGPNA转染胰腺癌Patu8988细胞的最高细胞结合率为(38.16±2.17)%,最高核内化率为(22.41±0.86)%。结论188Re直接标记经修饰的反基因肽核酸方法简便、标记率较高,能与胰腺癌细胞结合并转染入细胞核。

Objective To establish a stable method for labeling antigene peptide nucleic acid （AGPNA） with ^188Re. Methods The directly labeling method was adopted, and several labeling conditions were tested, such as the concentration of SnCl2.2H2O and the amount of AGPNA. The labeling efficiency was determined from 15min to 6 h after labeling. The in vitro stability of ^188ReAGPNA was analyzed by using human serum or sodium chloride as challenging agent, and the la- beling efficiency was determined from 15min to 24 h after added challenging agent. The total binding efficiency of ^188Re-AGPNA with pancreatic cancer cell Patu8988 and the bound of ^188ReAGPNA on the cellular nucleus was determined at various time points. Results The optimum labeling conditions were 100μl SnCl2·2H2O （20 mg/ml） and 20μl AGPNA （2 mg/ml）, incubated 30 min at room temperature. The labeling efficiency of ^188Re-AGPNA could reach （89.99±0.15）% and the amount of radiocolloid was （9.40±0.55）%. The labeling efficiency of ^188Re-AGPNA was （89.14±0.63）% after incubation for 24 h with human serum. The highest value of total binding efficiency of ^188Re-AGPNA with Patu8988 cell was （38.16±2.17）%, the highest binding value on the nucleus was increased to （22.41±0.86）% at the 24th hour. Conclusion This method of labeling AGPNA with ^188Re is stable and is high labeling efficiency ^188Re-AGPNA can bind with the pancreatic cancer cell Patu8988.