CAJ | 학술논문

目的：探讨E3泛素连接酶(EDD)基因在SKOV3/DDP细胞顺铂耐药中的作用。方法采用RT-PCR和Western blot检测人卵巢癌SKOV3细胞和SKOV3/DDP细胞内EDD表达水平的差异。通过转染EDD-siRNA沉默EDD的表达后,噻唑蓝法检测顺铂对SKOV3/DDP细胞增殖的影响,Annexin V-FITC/PI双染法检测顺铂对细胞凋亡的影响；Western blotting检测髓样细胞白血病-1(Mcl-1)和细胞周期检测点激酶2(CHK2)蛋白表达水平。结果 SKOV3细胞EDD的表达水平显著低于卵巢癌耐药细胞SKOV3/DDP (P<0.05)。 EDD-siRNA转染24 h后,EDD mRNA和蛋白表达水平均显著低于未转染组(P<0.05)；SKOV3/DDP细胞IC50值显著降低(P<0.05),Mcl-1蛋白的表达受到抑制(P<0.05),而CHK2无明显变化(P>0.05)。结论 EDD-siRNA能沉默EDD基因,降低Mcl-1蛋白的表达水平,增强顺铂对SKOV3/DDP细胞的促凋亡作用,降低SKOV3/DDP细胞对顺铂的耐药性,有效恢复SKOV3/DDP细胞顺铂敏感性。
목적：탐토E3범소련접매(EDD)기인재SKOV3/DDP세포순박내약중적작용。방법채용RT-PCR화Western blot검측인란소암SKOV3세포화SKOV3/DDP세포내EDD표체수평적차이。통과전염EDD-siRNA침묵EDD적표체후,새서람법검측순박대SKOV3/DDP세포증식적영향,Annexin V-FITC/PI쌍염법검측순박대세포조망적영향；Western blotting검측수양세포백혈병-1(Mcl-1)화세포주기검측점격매2(CHK2)단백표체수평。결과 SKOV3세포EDD적표체수평현저저우란소암내약세포SKOV3/DDP (P<0.05)。 EDD-siRNA전염24 h후,EDD mRNA화단백표체수평균현저저우미전염조(P<0.05)；SKOV3/DDP세포IC50치현저강저(P<0.05),Mcl-1단백적표체수도억제(P<0.05),이CHK2무명현변화(P>0.05)。결론 EDD-siRNA능침묵EDD기인,강저Mcl-1단백적표체수평,증강순박대SKOV3/DDP세포적촉조망작용,강저SKOV3/DDP세포대순박적내약성,유효회복SKOV3/DDP세포순박민감성。
Objective To discuss the effect of E3 isolated by differential Display (EDD) on Cisplatin resistance of SKOV3/DDP cells. Methods The expression difference of EDD between Human ovarian cancer cells SKOV3 and SKOV3/DDP cells was detected by Western blot and RT-PCR; EDD-siRNA was transfected into SKOV3/DDP cells to silence the expression of EDD, MTT assay was used to detect the chemosensitivity of SKOV3/DDP cells to Cisplatin, the apoptosis induced by Cisplatin was tested by Annexin V-FITC/PI, the protein levels of Mcl-1 and CHK2 was measured by Western blotting. Results The expression of EDD in SKOV3 cells was significantly lower than SKOV3/EDD cells (P<0.05). EDD expression was successful silenced and the IC50 of SKOV3/DDP cells was significantly decreased at 24 h in EDD-siRNA group compared with the control group (P< 0.05). Knockdown of EDD can significantly decrease IC50 and greatly down-regulate the expression of Mcl-1 (P< 0.05); however, EDD silence had no effect on CHK2 level (P>0.05). Conclusion Silencing EDD gene with EDD-siRNA reduce the expression of Mcl-1, enhance the apoptosis in-ducing effect of Cisplatin on SKOV3/DDP cells, depress the Cisplatin resistance of SKOV3/DDP cells and effectively recover SKOV3/DDP cell sensitivity to Cisplatin.