摘要
Abstract
[Objective] To explore whether antisense oligodeoxynucleotides(ASODN) targeting survivin gene can reverse apoptotic resistance induced by cisplatin in human lung adenocarcinoma cells. [Methods] A549/CDDP cell lines were cultured routinely with RPMI-1640 medium. Survivin ASODN mediated by cytofectin was transfected into the A549/CDDP cells .The influence of ASODN transfection on apoptosis was determined by Diphenylamine assay (DPA) ,caspase-3 colorimetric assay and flow cytometry. 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed to detect the cell viability, half-maximum inhibitory concentration (IC50) and cisplatin resistant index(RI) were thereby calculated, respectively. [Results] In the A549/CDDP cells transfected by survivin ASODN for 48 hrs, the percentage of DNA fragmentation, apoptotic index and caspase-3 activity was enhanced to (43.55±6.07)%, (34.03±2.25)% and (1.1298±0.2502),respectively (P ＜0.05), it was obviously significant,compared to the controls. While combination with ASODN and 10 μmol/L cisplatin caused far more obvious apoptotic alterations in the cells, of which the percentage of DNA fragmentation, AI and caspase-3 activity was reached to (76.73±2.04)%, (65.85±5.47)% and (1.6805±0.2758)(P ＜0.05), respectively, and even compared to single ASODN group, it was still significant (P ＜0.05). Transfected with survivin ASODN single and with combination of cisplatin for 48 hrs, the rate of cell growth inhibition in the resistant cells was enhanced to 59.3% and 83.7%, respectively(P＜0.05), while cell viability inversely had the lowest reduction. And the half-maximum inhibitory concentration of cisplatin was reduced from (225.03±10.59)μmol/L to(158.84±4.256) μmol/L, while the resistant index was conversely reduced from 11.9 to 8.39. Non-sense oligodeoxynucleotides(NSODN) and liposome had no effect on the cells(P＞0.05). [Conclusion] Transfection of antisense oligodeoxynucleotides targeting survivin gene can reduce the apoptosis threshold, thereafter to great extent reverse apoptotic resistance induced by cisplatin in cisplatin-resistant human lung adenocarcinoma in vitro.关键词
survivin基因/反义寡核苷酸/多药耐药性/细胞凋亡Key words
survivin gene/antisense oligodeoxynucleotides/multidrug resistance/apoptosis分类
医药卫生
