中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
5期
1072-1074
,共3页
微小RNA-21%食管癌%顺铂
微小RNA-21%食管癌%順鉑
미소RNA-21%식관암%순박
MicroRNA-21%Esophageal cancer%Cisplatin
目的 观察微小RNA-21(miRNA-21)反义寡核苷酸对食管癌EC9706顺铂(DDP)耐药细胞(EC9706/DDP)的影响.方法 设计miRNA-21反义寡核苷酸,建立EC9706/DDP耐药细胞株并转染耐药细胞,实时荧光定量聚合酶链反应(FQ-PCR)检测各组细胞miRNA-21表达水平,噻唑蓝(MTT)法检测各组细胞对DDP药物敏感性、克隆形成实验检测各组细胞增殖能力,流式细胞术检测细胞凋亡变化,Western blot检测B细胞淋巴瘤/白血病-2(bcl-2)、B细胞淋巴瘤/白血病-2相关X蛋白(bax)蛋白表达.结果 与空白组、miRNA-阴性对照(NC)比较,miRNA-21反义寡核苷酸组miRNA-21相对表达量下降为0.21 ±0.89、半数抑制剂量(IC50)明显降低为(16.22 ±1.85) mg/L、克隆集落的数量较少、细胞凋亡率明显增高为(30.5±2.9)％、bcl-2蛋白表达水平降低,bax蛋白表达水平增高,差异均有统计学意义(P＜0.05).结论 miRNA-21可能通过降低bcl-2、增高bax蛋白表达水平降低EC9706/DDP细胞对DDP的耐药性.
目的 觀察微小RNA-21(miRNA-21)反義寡覈苷痠對食管癌EC9706順鉑(DDP)耐藥細胞(EC9706/DDP)的影響.方法 設計miRNA-21反義寡覈苷痠,建立EC9706/DDP耐藥細胞株併轉染耐藥細胞,實時熒光定量聚閤酶鏈反應(FQ-PCR)檢測各組細胞miRNA-21錶達水平,噻唑藍(MTT)法檢測各組細胞對DDP藥物敏感性、剋隆形成實驗檢測各組細胞增殖能力,流式細胞術檢測細胞凋亡變化,Western blot檢測B細胞淋巴瘤/白血病-2(bcl-2)、B細胞淋巴瘤/白血病-2相關X蛋白(bax)蛋白錶達.結果 與空白組、miRNA-陰性對照(NC)比較,miRNA-21反義寡覈苷痠組miRNA-21相對錶達量下降為0.21 ±0.89、半數抑製劑量(IC50)明顯降低為(16.22 ±1.85) mg/L、剋隆集落的數量較少、細胞凋亡率明顯增高為(30.5±2.9)％、bcl-2蛋白錶達水平降低,bax蛋白錶達水平增高,差異均有統計學意義(P＜0.05).結論 miRNA-21可能通過降低bcl-2、增高bax蛋白錶達水平降低EC9706/DDP細胞對DDP的耐藥性.
목적 관찰미소RNA-21(miRNA-21)반의과핵감산대식관암EC9706순박(DDP)내약세포(EC9706/DDP)적영향.방법 설계miRNA-21반의과핵감산,건립EC9706/DDP내약세포주병전염내약세포,실시형광정량취합매련반응(FQ-PCR)검측각조세포miRNA-21표체수평,새서람(MTT)법검측각조세포대DDP약물민감성、극륭형성실험검측각조세포증식능력,류식세포술검측세포조망변화,Western blot검측B세포림파류/백혈병-2(bcl-2)、B세포림파류/백혈병-2상관X단백(bax)단백표체.결과 여공백조、miRNA-음성대조(NC)비교,miRNA-21반의과핵감산조miRNA-21상대표체량하강위0.21 ±0.89、반수억제제량(IC50)명현강저위(16.22 ±1.85) mg/L、극륭집락적수량교소、세포조망솔명현증고위(30.5±2.9)％、bcl-2단백표체수평강저,bax단백표체수평증고,차이균유통계학의의(P＜0.05).결론 miRNA-21가능통과강저bcl-2、증고bax단백표체수평강저EC9706/DDP세포대DDP적내약성.
Objective To investigate the impact of microRNA (miRNA)-21 antisense oligonucleotide on cisplatin (DDP) resistance of EC9706/DDP cells.Methods miRNA-21 antisense oligonucleotides were designed and transfected into EC9706/DDP-resistant cells.The miRNA-21 expression levels were detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).Methyl thiazol tetrazolium (MTT) method was used to measure the sensitivity of cells to DDP drugs.Colony formation assay was used to test the cell proliferation.The change of apoptosis was examined by flow cytometry.Western blotting was performed to detect the expression of B cell lymphoma/leukemia-2 (bcl-2),and bcl-2 associated X protein (bax) proteins.Results As compared with miRNA-NC negative control group and blank control group,the miRNA-21 relative expression was decreased to (0.21 ±0.89),50％ inhibitory dose (IC50) decreased to (16.22 ± 1.85) mg/L,the number of clonal colonies was reduced,the apoptosis rate significantly increased to (30.5 ± 2.9) ％,bcl-2 protein expression decreased,and bax protein expression increased in miRNA-21 antisense oligonucleotide group (P ＜ 0.05).Conclusion miRNA-21 decreased the resistance of EC9706/DDP cells to DDP possibly through reducing the bcl-2 protein expression,and increasing the bax protein expression.
