CAJ | 학술논문

目的：研究小干扰核糖核酸（siRNA）沉默多药耐药相关蛋白（MRP）3基因（MRP3-siRNA）对原发性肝癌（肝癌）细胞耐药性的影响。方法应用不同梯度浓度的多柔比星（ADM）制备肝癌耐药细胞株HepG2/ADM，用携带MRP3-siRNA的lipofectamine 2000脂质体转染HepG2/ADM细胞。分为HepG2细胞组（对照组）、HepG2/ADM细胞组（耐药组）和转染MRP3-siRNA的HepG2/ADM细胞组（干扰组）。采用实时荧光定量聚合酶链反应检测3组细胞MRP3基因信使核糖核酸（mRNA）含量，用蛋白质印迹法检测MRP3蛋白表达，噻唑蓝法检测肝癌细胞对ADM、氟尿嘧啶（5-FU）、长春新碱、奥沙利铂的半数抑制浓度（IC50），并计算耐药指数（RI）。3组实验数据比较采用单因素方差分析，两两比较采用LSD-t 或t检验。结果耐药组的MRP3 mRNA平均含量（5.16±0.31）较对照组（3.08±0.27）和干扰组（2.85±0.23）明显升高（LSD-t=8.765，10.363；P<0.05）。耐药组的MRP3蛋白含量（21063±274）较对照组（14476±217）和干扰组（6660±153）明显升高（LSD-t=21.836，79.578；P<0.05）。耐药组对ADM、5-FU、长春新碱、奥沙利铂的RI分别为14.40±0.31、26.68±0.22、28.70±0.49、20.23±0.54，干扰组相应为3.55±0.16、9.60±0.27、2.11±0.17、3.15±0.13，干扰组的RI明显低于耐药组（t=53.873，84.933，88.811，53.258；P<0.05）。结论 siRNA沉默肝癌细胞MRP3基因可提高肝癌细胞对化疗药物的敏感性，逆转肝癌细胞对化疗药物的耐药性。
목적：연구소간우핵당핵산（siRNA）침묵다약내약상관단백（MRP）3기인（MRP3-siRNA）대원발성간암（간암）세포내약성적영향。방법응용불동제도농도적다유비성（ADM）제비간암내약세포주HepG2/ADM，용휴대MRP3-siRNA적lipofectamine 2000지질체전염HepG2/ADM세포。분위HepG2세포조（대조조）、HepG2/ADM세포조（내약조）화전염MRP3-siRNA적HepG2/ADM세포조（간우조）。채용실시형광정량취합매련반응검측3조세포MRP3기인신사핵당핵산（mRNA）함량，용단백질인적법검측MRP3단백표체，새서람법검측간암세포대ADM、불뇨밀정（5-FU）、장춘신감、오사리박적반수억제농도（IC50），병계산내약지수（RI）。3조실험수거비교채용단인소방차분석，량량비교채용LSD-t 혹t검험。결과내약조적MRP3 mRNA평균함량（5.16±0.31）교대조조（3.08±0.27）화간우조（2.85±0.23）명현승고（LSD-t=8.765，10.363；P<0.05）。내약조적MRP3단백함량（21063±274）교대조조（14476±217）화간우조（6660±153）명현승고（LSD-t=21.836，79.578；P<0.05）。내약조대ADM、5-FU、장춘신감、오사리박적RI분별위14.40±0.31、26.68±0.22、28.70±0.49、20.23±0.54，간우조상응위3.55±0.16、9.60±0.27、2.11±0.17、3.15±0.13，간우조적RI명현저우내약조（t=53.873，84.933，88.811，53.258；P<0.05）。결론 siRNA침묵간암세포MRP3기인가제고간암세포대화료약물적민감성，역전간암세포대화료약물적내약성。
Objective To observe the impact of multidrug resistance-related protein (MRP) 3 gene silenced by small interfering RNA (siRNA) on the drug resistance of primary liver cancer cells. Methods The drug resistance cell lines HepG2/Adriamycin(ADM) were developed by exposing parental cells to stepwise increasing concentrations of ADM and then MRP3-siRNAs were transfected in HepG2/ADM cells with lipofectamine 2000 liposomes. Three groups were assigned:HepG2 cell group (control group), HepG2/ADM cell group (resistance group) and MRP3-siRNA transfected HepG2/ADM cell group (interference group). The MRP3 mRNA contents of 3 groups were measured by real-time fluorescent quantitative polymerase chain reaction (PCR). The expression of MRP3 protein was detected by Western blot. The 50%inhibitory concentrations (IC50) of ADM, lfuorouracil (5-FU), vincristine, oxaliplatin were determined by methyl thiazolyl tetrazolium (MTT) and the drug resistance indexes (RI) were calculated. The experimental data of 3 groups were compared by one-way analysis of variance, while pairwise comparisons were conducted using LSD-t or t test. Results The mean of MRP3 mRNA content in resistance group (5.16±0.31) was signiifcantly higher than those in control group (3.08±0.27) and interference group (2.85±0.23) (LSD-t=8.765, 10.363;P<0.05). The MRP3 protein content in resistance group (21 063±274) was signiifcantly higher than those in control group (14 476±217) and interference group (6 660±153) (LSD-t=21.836, 79.578;P<0.05). The RI of ADM, 5-FU,vincristine, oxaliplatin in resistance group were 14.40±0.31, 26.68±0.22, 28.70±0.49, 20.23±0.54, and were 3.55±0.16, 9.60±0.27, 2.11±0.17, 3.15±0.13 respectively in interference group. The RI in interference group were signiifcantly lower than those in resistance group (t=53.873, 84.933, 88.811, 53.258; P<0.05). Conclusions MRP3 gene of liver cancer cells silenced by siRNA can improve the cells' sensitivity to chemotherapy drugs and reverse its chemotherapy drug resistance.