Survivin of Antisense Oligodeoxynucleotides Reverse Apoptotic Resistance Induced by Cisplatin in Lung Cells

摘要

We explore whether antisense oligodeoxy-nucleotides (ASODN) targeting survivin gene can reverse apoptotic resistance induced by cisplatin in human lung adenocar-cinoma cells．A549／CDDP cell lines were cultured routinely with RPMI-1640 medium．Survivin ASODN mediated by cytofectin was transfeeted 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-dimethyl-thiazol-2-y1)-2，5-diphenyl tetra-zolium bromide(MTT)assay was performed to detect the cel viabihty，half-maximum inhibitory concentration OC,o)and cisplatin resistant index were thereby calculated，respectively. In the A549／CDDP cells transfected by survivin ASODN for 48 hrs，the percentage of DNA fragmen-tation，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 witIl ASODN and 10 bmol／L cisplatin caused far more obvious apoptotic altera-tions in the cels，ofwhich the percentage of DNA fragm entation，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 comb ination of cisplatin f0r 48 hrs，the rate of cel growth inhibition in the resistant cels was enhanced to 59.3％and 83.7％,respe- - ctively(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 l1.9 to 8.39．Non-sense oligodeoxynucleotides (NSODN1 and liposome had no efect on the cells(p>0.05). Transfection of antisense oligodeoxynucleotides targeting survivin gene can reduce the apop-tosis threshold，thereafter to great extent reverse apoptotic resis-tance induced by cisplatin in cisplatin-resistant human lung adenocarcinoma in vitro.