【摘要】 目的:研究反义硫代磷酸化寡核苷酸(phosphorothioate antisense o1igonuc1eotide,PS-ASOs)抑制人舌癌细胞耐放疗株Tca8113多药耐药基因1(multidrug resistance gene1,MDR1)以及MDR1蛋白(P-gp)表达的作用。方法:人工合成互补于MDR1基因mRNA特定片断的硫代磷酸化正反义寡核苷酸;PS-ASOs浓度分别为0.1μmol/L,0.25μmol/L,0.5μmol/L,以正义寡核苷酸作为实验对照,未转染的耐放疗组为空白对照,以5μl/ml脂质体Lipofectamine为载体转染耐放疗的Tca8113细胞株;逆转录多聚酶链反应(RT-PCR)测定转染后MDR1 mRNA表达,流式细胞技术(FCM)测定细胞膜P-gp表达。结果:与正义组和空白对照组相比,转染PS-ASOs后12h,MDR1 mRNA和P-gp表达降低,转染24h,各剂量组表达均降至最低,在转染48h后,基本恢复到转染前水平,而正义组和空白对照组在各时段及各剂量组均无统计学差异;双因素方差分析:PS-ASOs不同浓度(P=0.00)、不同作用时间(P=0.00)对下调MDR1 mRNA、P-gp表达差异显著,结论:PS-ASOs能降低MDR1基因以及蛋白的表达;PS-ASOs的作用具有浓度依赖性和时间依赖性。更多还原

【Abstract】 Objective:To study the efficacy of phosphrothioate antisense oligonucleotides(PS-ASOs)on suppressing the expression of Multidrug Resistance Gene 1(MDR1)and its protein(P-glycoprotein,P-gp)in radiation-resistance hu-man tongue carcinoma cell line(RR-Tca8113).Method:RR-Tca8113 Cell line was transfected with 0.1 μmol/L,0.25 μmol/L,0.5 μmol/L synthetic PS-ASOs complementing to the coding region of MDR1 mRNA with 5 μL/mL Lipofec-tamine as a carrier.The sense SOs(PS-SOs)tranfection cell and no transfection cell were used as controls.The expression of MDR1 mRNA and P-gp was detected by reverse transcription-polymerase chain reaction(RT-PCR)and flow cytometry(FCM)in 12 h,24 h,and 48 h after transfection,respectively.Result:The MDR1 mRNA and P-gp expression were no statistic difference between PS-SOs transfection groups and no transfection groups in every time group.Comparing with PS-SOs and no transfection groups,the expression of MDR1 mRNA and P-gp of PS-ASOs group was declined in 12 h,to the lowest in 24 h,and recovered in 48 h in every concentration group.On the other hand,as the decrease of PS-ASOs concen-tration,the downregulate effect of MDR1 expression was decrease.There were significantly statistic difference among differ-ent concentrations(P=0.00)and different times(P=0.00)to downregulate the expression of MDR1 mRNA and P-gp.Con-clusion:PS-ASOs reduces the expression of MDR1 mRNA and P-gp of human tongue carcinoma cell line with concentra-tion and time dependence.更多还原