Survivin of Antisense 0ligodeoxynucleotides Reverse Apoptotic Resistance Induced by Cisplatin in Lung Cells
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 ％ ,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 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.
Zhi-wei Zhang Dong Jiang Jin-feng Ge Shi-ying Zheng
Department of Thoracocardiac Surgery, The First Affiliated Hospital of Suzhou University,Suzhou,Jiangsu, 215006, China
国际会议
成都
英文
1-6
2010-06-18（万方平台首次上网日期，不代表论文的发表时间）