中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
18期
1422-1426
,共5页
周卫%赵梓彤%刘玲燕%詹启敏%宋咏梅
週衛%趙梓彤%劉玲燕%詹啟敏%宋詠梅
주위%조재동%류령연%첨계민%송영매
肺肿瘤%紫杉醇%抗药性,肿瘤%细胞凋亡%适体,核苷
肺腫瘤%紫杉醇%抗藥性,腫瘤%細胞凋亡%適體,覈苷
폐종류%자삼순%항약성,종류%세포조망%괄체,핵감
Lung neoplasms%Taxol%Drug resistance,neoplasm%Apoptosis%Aptamers,nucleotide
目的 探讨核酸适配子AS1411对紫杉醇耐药肺腺癌A549细胞(A549/T细胞)凋亡的影响.方法 采用0 ～ 20.0 μmol/L浓度的AS1411处理A549/T细胞,甲基甲苯基硫细胞检测(MTS)实验、平板克隆形成实验检测细胞活性[吸光度值(A490nm)]及增殖能力变化,流式细胞仪检测对细胞凋亡的影响,同时应用蛋白免疫印迹法检测凋亡信号通路相关蛋白表达的变化.结果 A549/T细胞呈现部分上皮细胞间质化(EMT)、表皮生长因子受体(EGFR)表达缺失等特征.经5.0 μmol/L的AS1411处理后,与对照序列相比,A549/T细胞的细胞活性显著降低(A490nm:0.185±0.009比0.272±0.006,P＜0.001)、克隆形成数显著减少(74±13比120±12,P=0.010);随AS1411浓度的增加,细胞活性明显降低,呈剂量依赖性.用20.0 μmol/L的AS1411处理48 h后,A549/T细胞凋亡率显著高于对照序列[(19.9±2.6)％比(8.8±1.3)％,P=0.002],同时蛋白激酶B(AKT)、细胞外信号调节激酶1/2(ERK1/2)和B细胞淋巴瘤因子2(Bcl-2)表达均显著低于对照序列处理组(0.353±0.003、0.432±0.015、0.294 ±0.015比0.688±0.003、0.911±0.019、0.422±0.018,均P＜0.001).结论 AS1411可通过抑制AKT-ERK通路而促进A549/T细胞凋亡.
目的 探討覈痠適配子AS1411對紫杉醇耐藥肺腺癌A549細胞(A549/T細胞)凋亡的影響.方法 採用0 ～ 20.0 μmol/L濃度的AS1411處理A549/T細胞,甲基甲苯基硫細胞檢測(MTS)實驗、平闆剋隆形成實驗檢測細胞活性[吸光度值(A490nm)]及增殖能力變化,流式細胞儀檢測對細胞凋亡的影響,同時應用蛋白免疫印跡法檢測凋亡信號通路相關蛋白錶達的變化.結果 A549/T細胞呈現部分上皮細胞間質化(EMT)、錶皮生長因子受體(EGFR)錶達缺失等特徵.經5.0 μmol/L的AS1411處理後,與對照序列相比,A549/T細胞的細胞活性顯著降低(A490nm:0.185±0.009比0.272±0.006,P＜0.001)、剋隆形成數顯著減少(74±13比120±12,P=0.010);隨AS1411濃度的增加,細胞活性明顯降低,呈劑量依賴性.用20.0 μmol/L的AS1411處理48 h後,A549/T細胞凋亡率顯著高于對照序列[(19.9±2.6)％比(8.8±1.3)％,P=0.002],同時蛋白激酶B(AKT)、細胞外信號調節激酶1/2(ERK1/2)和B細胞淋巴瘤因子2(Bcl-2)錶達均顯著低于對照序列處理組(0.353±0.003、0.432±0.015、0.294 ±0.015比0.688±0.003、0.911±0.019、0.422±0.018,均P＜0.001).結論 AS1411可通過抑製AKT-ERK通路而促進A549/T細胞凋亡.
목적 탐토핵산괄배자AS1411대자삼순내약폐선암A549세포(A549/T세포)조망적영향.방법 채용0 ～ 20.0 μmol/L농도적AS1411처리A549/T세포,갑기갑분기류세포검측(MTS)실험、평판극륭형성실험검측세포활성[흡광도치(A490nm)]급증식능력변화,류식세포의검측대세포조망적영향,동시응용단백면역인적법검측조망신호통로상관단백표체적변화.결과 A549/T세포정현부분상피세포간질화(EMT)、표피생장인자수체(EGFR)표체결실등특정.경5.0 μmol/L적AS1411처리후,여대조서렬상비,A549/T세포적세포활성현저강저(A490nm:0.185±0.009비0.272±0.006,P＜0.001)、극륭형성수현저감소(74±13비120±12,P=0.010);수AS1411농도적증가,세포활성명현강저,정제량의뢰성.용20.0 μmol/L적AS1411처리48 h후,A549/T세포조망솔현저고우대조서렬[(19.9±2.6)％비(8.8±1.3)％,P=0.002],동시단백격매B(AKT)、세포외신호조절격매1/2(ERK1/2)화B세포림파류인자2(Bcl-2)표체균현저저우대조서렬처리조(0.353±0.003、0.432±0.015、0.294 ±0.015비0.688±0.003、0.911±0.019、0.422±0.018,균P＜0.001).결론 AS1411가통과억제AKT-ERK통로이촉진A549/T세포조망.
Objective To explore the effects of AS1411 on the apoptosis of taxol-resistant lung adenocarinoma A549 cell (A549/T cell).Methods A549/T cells were treated with AS1411 at a concentration gradient of 0-20.0 μmol/L.The assays of methyl tolyl sulfide (MTS) and colony formation were used to detect the cellular vitality (absorbance value (A490,nm)) and proliferation.The apoptotic effects were detected by flow cytometer and the relevant apoptotic signaling proteins detected by Western blot.Results A549/T cells exhibited some characteristics of epithelial mesenchymal transition (EMT) and a negative expression of epidermal growth factor receptor (EGFR).After a treatment of 5.0 μmol/L AS1411,compared to the control sequence,cell vitality was inhibited (A490 nm:0.185 ± 0.009 vs 0.272 ± 0.006,P ＜0.001) and the number of clone formation decreased (74 ± 13 vs 120 ± 12,P =0.010).With rising AS1411 concentration,A549/T cells vitality decreased in a dose-dependent manner.After a 48-hour treatment of 20.0 μmol/L AS1411,the ratio of apoptosis ((19.9 ± 2.6)％) had significant difference (P =0.002) with the control sequence group ((8.8 ± 1.3) ％).Compared to the control sequence group,the expressions of protein kinase B (AKT),extracellular regulated protein kinases 1/2 (ERK1/2) and B-cell lymphoma 2 (Bcl-2) protein declined (0.353 ± 0.003,0.432 ± 0.015,0.294 ± 0.015 vs 0.688 ±0.003,0.911 ± 0.019,0.422 + 0.018,all P ＜ 0.001).Conclusion AS1411 may induce the apoptosis of A549/T cells through inhibiting the AKT-ERK pathways.
