CAJ | 학술논문

目的：探讨抑制microRNA-192（miR-192）在非小细胞肺癌A549/DDP细胞对顺铂（DDP）耐药性方面的影响及可能机制。方法通过实时荧光定量PCR（ qRT-PCR）法检测人肺腺癌耐DDP细胞株A549/DDP及其亲本细胞株A549细胞中miR-192的表达水平，A549/DDP细胞转染miR-192抑制剂（inhibitor）和miR-阴性对照（NC）48h后采用qRT-PCR检测转染效率，分别采用MTT法、克隆形成实验及流式细胞术检测转染48h后A549/DDP细胞对DDP的药物敏感性、细胞增殖能力及细胞凋亡变化，Western blotting检测转染48h后细胞中Bax和Bcl-2的表达变化。结果 miR-192在A549/DDP细胞中的表达水平高于A549细胞（ P＜0?05）；转染 miR-192 inhibitor 48h 后的 A549/DDP 细胞 miR-192水平低于转染 miR-NC 者（ P＜0?05）；转染miR-192 inhibitor后，A549/DDP细胞的增殖能力减弱、凋亡细胞增多、DDP对其半数抑制浓度降低、Bax蛋白水平升高和Bcl-2蛋白水平下降，与转染miR-NC者比较，差异均有统计学意义（ P＜0?05）。结论抑制miR-192能够降低A549/DDP细胞对DDP的耐药性，其作用机制可能是通过增加细胞凋亡以及下调Bcl-2蛋白和上调Bax蛋白表达来实现的。
목적：탐토억제microRNA-192（miR-192）재비소세포폐암A549/DDP세포대순박（DDP）내약성방면적영향급가능궤제。방법통과실시형광정량PCR（ qRT-PCR）법검측인폐선암내DDP세포주A549/DDP급기친본세포주A549세포중miR-192적표체수평，A549/DDP세포전염miR-192억제제（inhibitor）화miR-음성대조（NC）48h후채용qRT-PCR검측전염효솔，분별채용MTT법、극륭형성실험급류식세포술검측전염48h후A549/DDP세포대DDP적약물민감성、세포증식능력급세포조망변화，Western blotting검측전염48h후세포중Bax화Bcl-2적표체변화。결과 miR-192재A549/DDP세포중적표체수평고우A549세포（ P＜0?05）；전염 miR-192 inhibitor 48h 후적 A549/DDP 세포 miR-192수평저우전염 miR-NC 자（ P＜0?05）；전염miR-192 inhibitor후，A549/DDP세포적증식능력감약、조망세포증다、DDP대기반수억제농도강저、Bax단백수평승고화Bcl-2단백수평하강，여전염miR-NC자비교，차이균유통계학의의（ P＜0?05）。결론억제miR-192능구강저A549/DDP세포대DDP적내약성，기작용궤제가능시통과증가세포조망이급하조Bcl-2단백화상조Bax단백표체래실현적。
Objective To explore the effect of microRNA-192(miR-192) in enhancing cisplatin(DDP) resistance of non-small cell lung(NSCLC) A549/DDP cell line and elucidate its related mechanism. Methods The real-time quantitative PCR(qRT-PCR) was used to detect miR-192 expression in A549 cells and A549/DDP cells. The A549/DDP cells were transfected with miR-192 inhibitor or miR-negative control(NC) and qRT-PCR was used to test the transfection efficiency. Drug sensitivity to DDP 48h after transfection was tested by MTT assay. Colony formation assay was used to detect the proliferation of A549/DDP cells 48h after transfec-tion. Flow cytometric analysis was used to observe apoptosis of A549/DDP cells 48h after transfection on treatment with DDP. The pro-tein expressions of Bax and Bcl-2 in A549/DDP cells after transfection were detected by Western blotting method. Results The miR-192 expression in A549/DDP cell line was significantly higher than that in A549 cell line( P<0?05) . The miR-192 expression was sig-nificantly decreased at 48h after transfection of miR-192 inhibitor than that of miR-NC( P<0?05) . The half inhibition concentration of DDP was significantly lower in A549/DDP cells transfection of miR-192 inhibitor than miR-NC ( P<0?05 ) . Colony formation assay showed that the proliferation of miR-192 inhibitor transfected A549/DDP cells was significantly inhibited(P<0?05). Compared with miR-NC A549/DDP cells, the apoptosis of cells transfected with miR-192 inhibitor increased after treatment with DDP(P<0?05). Fur-thermore, the down-regulated Bcl-2 and up-regulated Bax were observed in A549/DDP cells transfection of miR-192 inhibitor versus miR-NC( P<0?05) . Conclusion Inhibiting expression of miR-192 could reduce the resistance of A549/DDP cells to DDP with the possible mechanism of inducing apoptosis, down-regulating the expressions of Bcl-2 protein and up-regulating the expression of Bax pro-tein.