论文部分内容阅读
目的探讨利用脂质体转染125I标记的CerbB-2反义肽核酸(asPNA)阻断人卵巢癌细胞株SKOV3中CerbB-2mRNA翻译并抑制Her-2蛋白表达的作用。方法 (1)125I用CH-T法标记CerbB-2-asPNA,测定其标记率和放化纯。(2)分对照组、非标记分子探针组和125I标记反义肽核酸组,运用RT-PCR法检测各组细胞CerbB-2mRNA的翻译。(3)运用流式细胞仪检测各组细胞的Her-2蛋白表达率。结果 (1)125I标记CerbB-2-asPNA的标记率为(56.90±4.38)%,放射化学纯度为(87.00±0.99)%,放射性浓度为3.7MBq/ml,化学浓度为5.00μmol/L。(2)125I标记CerbB-2-asPNA作用SKOV3细胞24h后,SKOV3细胞CerbB-2在mRNA水平上明显下降,与SKOV3细胞对照组比较,差异有统计学意义(P<0.01)。(3)125I标记CerbB-2-asPNA作用SKOV3细胞24h后,细胞的Her-2蛋白表达率降低幅度较大,与SKOV3细胞组对比,两者间差异有统计学意义(P<0.01)。结论 125I-L-CerbB-2-asP-NA对人卵巢癌细胞株SKOV3CerbB-2基因表达在mRNA翻译和蛋白表达水平上均有抑制作用。
Objective To investigate the effect of liposome-transfected CerbB-2 antisense oligodeoxynucleotide (asPNA) on the translation of CerbB-2 mRNA and the inhibition of Her-2 protein expression in human ovarian cancer cell line SKOV3. Method (1) 125I CerbB-2-asPNA was labeled with CH-T method and its labeling rate and radiochemical purity were determined. (2) The control group, the unlabeled molecular probe group and the 125I labeled antisense peptide nucleic acid group were used to detect the translation of CerbB-2 mRNA in each group by RT-PCR. (3) The expression of Her-2 protein in each group of cells was detected by flow cytometry. Results (1) The labeling rate of 125I-labeled CerbB-2-asPNA was (56.90 ± 4.38)%, the radiochemical purity was 87.00 ± 0.99%, the radioactive concentration was 3.7MBq / ml and the chemical concentration was 5.00μmol / L. (2) CerbB-2 of SKOV3 cells was significantly decreased after treated with 125I-labeled CerbB-2-asPNA for 24 hours. Compared with the SKOV3 cells, the difference was statistically significant (P <0.01). (3) The expression of Her-2 in SKOV3 cells treated with 125I-labeled CerbB-2-asPNA for 24 h reduced significantly, compared with that of SKOV3 cells (P <0.01). Conclusion 125I-L-CerbB-2-asP-NA can inhibit the mRNA translation and protein expression of SKOV3CerbB-2 in human ovarian cancer cell line.