CAJ | 학술논문

目的：探讨c-FLIP siRNA干扰c-FLIP mRNA水平对阿霉素耐药细胞K562/ADR的耐药性的影响及作用机制。方法应用siRNA干扰的方法抑制K562及K562/ADR细胞c-FLIP的表达，通过荧光定量PCR的方法检测c-FLIP mRNA表达及对多药耐药基因MDR1 mRNA水平的影响。MTT法检测c-FLIP干扰与否对K562及K562/ADR细胞增殖的影响，Annexin V/7-ADD双染研究c-FLIP干扰与否对K562及K562/ADR细胞凋亡的影响。结果与阴性siRNA转染组相比较，c-FLIP siRNA转染下调K562细胞中c-FLIP的mRNA后，K562细胞增殖受到一定程度的抑制(P<0.05)，但是并未显著诱导细胞凋亡，c-FLIP干扰与否K562细胞48 h增殖率分别为(69.14±1.82)%和(60.69±2.23)%，凋亡率分别为(1.7±0.3)%和(1.8±0.2)%。与阴性siRNA转染组相比较， c-FLIP siRNA转染抑制K562/ADR细胞中c-FLIP的mRNA后，K562/ADR细胞增殖显著被抑制(P<0.05)，并显著诱导了细胞凋亡增加(P<0.05)，c-FLIP干扰与否K562/ADR细胞48 h增殖率分别为(-6.07±0.71)%和(-37.45±3.53)%，凋亡率分别为(5.2±0.4)%和(9.2±0.4)%。并且c-FLIP siRNA下调c-FLIP mRNA水平后，K562/ADR细胞中的多药耐药基因MDR1的mRNA表达水平也被显著下调(P<0.05)。结论 c-FLIP siRNA下调K562/ADR细胞中c-FLIP的mRNA水平抑制了多药耐药基因MDR1的表达，从而抑制了K562/ADR细胞对阿霉素的耐药性。
목적：탐토c-FLIP siRNA간우c-FLIP mRNA수평대아매소내약세포K562/ADR적내약성적영향급작용궤제。방법응용siRNA간우적방법억제K562급K562/ADR세포c-FLIP적표체，통과형광정량PCR적방법검측c-FLIP mRNA표체급대다약내약기인MDR1 mRNA수평적영향。MTT법검측c-FLIP간우여부대K562급K562/ADR세포증식적영향，Annexin V/7-ADD쌍염연구c-FLIP간우여부대K562급K562/ADR세포조망적영향。결과여음성siRNA전염조상비교，c-FLIP siRNA전염하조K562세포중c-FLIP적mRNA후，K562세포증식수도일정정도적억제(P<0.05)，단시병미현저유도세포조망，c-FLIP간우여부K562세포48 h증식솔분별위(69.14±1.82)%화(60.69±2.23)%，조망솔분별위(1.7±0.3)%화(1.8±0.2)%。여음성siRNA전염조상비교， c-FLIP siRNA전염억제K562/ADR세포중c-FLIP적mRNA후，K562/ADR세포증식현저피억제(P<0.05)，병현저유도료세포조망증가(P<0.05)，c-FLIP간우여부K562/ADR세포48 h증식솔분별위(-6.07±0.71)%화(-37.45±3.53)%，조망솔분별위(5.2±0.4)%화(9.2±0.4)%。병차c-FLIP siRNA하조c-FLIP mRNA수평후，K562/ADR세포중적다약내약기인MDR1적mRNA표체수평야피현저하조(P<0.05)。결론 c-FLIP siRNA하조K562/ADR세포중c-FLIP적mRNA수평억제료다약내약기인MDR1적표체，종이억제료K562/ADR세포대아매소적내약성。
Objective To investigate the effect of silenced c-FLIP mRNA level by small interfering RNA (siRNA) on adriamycin-resistance of K562/ADR cells. Methods c-FLIP siRNA and negative siRNA were transfect-ed into K562 and K562/ADR cell lines respectively, and mRNA expression of c-FLIP and multi-drug resistance gene 1 (MDR1) were detected by quantitative PCR. Cell proliferation rate was detected by MTT assay, and cell apoptosis rate was assayed by Annexin V/7-ADD double-staining method. Results Compared with negative siRNA transfection group, siRNA transfection significantly decreased c-FLIP mRNA expression in K562 cells (P<0.05) and slightly inhib-ited the proliferation of K562 cell (not enough to induce K562 cell apoptosis). The 48 h proliferation rates of K562 cells were (69.14 ± 1.82)% and (60.69 ± 2.23)% in negative siRNA transfection group and siRNA transfection group, and the apoptosis rate were (1.7±0.3)%and (1.8±0.2)%, respectively. c-FLIP siRNA transfection significantly inhibit-ed K562/ADR proliferation and induced cell apoptosis (P<0.05). The 48 h proliferation rate of K562/ADR cells were (-6.07 ± 0.71)% and (-37.45 ± 3.53)% in negative siRNA transfection group and siRNA transfection group, and the apoptosis rate were (5.2 ± 0.4)% and (9.2 ± 0.4)%, respectively. In addition, MDR1 mRNA expression of K562/ADR was significantly decreased (P<0.05) upon c-FLIP siRNA transfection (P<0.05). Conclusion c-FLIP siRNA transfec-tion significantly decreases mRNA expression of MDR1 and inhibites the adriamycin-resistance of K562/ADR cells.