CAJ | 학술논문

目的：研究小干扰RNA（siRNA）对预分化骨髓间充质干细胞（BMSCs）β2-微球蛋白（β2M）基因表达的影响。方法：BMSCs成软骨诱导液预分化21 d后，将β2 M siRNA用Lipofectamine 2000转染BMSCs、分为转染组、空白对照组和阴性对照组，应用实时定量PCR、Western blotting、激光共聚焦显微观察等方法检测siRNA对预分化BMSCs的β2 M mRNA和蛋白的表达情况，甲苯胺蓝和Ⅱ型胶原免疫荧光检测预分化BMSC的蛋白聚糖多聚体和Ⅱ型胶原分泌情况。结果：实时定量PCR、Western blotting 和激光共聚焦显微观察结果显示siRNA成功抑制β2 M mRNA和蛋白的表达，甲苯胺蓝和Ⅱ型胶原免疫荧光结果显示siRNA不影响预分化BMSCs蛋白聚糖多聚体和Ⅱ型胶原蛋白的分泌。结论： RNAi 干扰β2 M 可降低BMSCs 中β2 M mRNA和蛋白的表达，但不影响预分化BMSCs的软骨细胞特性。
목적：연구소간우RNA（siRNA）대예분화골수간충질간세포（BMSCs）β2-미구단백（β2M）기인표체적영향。방법：BMSCs성연골유도액예분화21 d후，장β2 M siRNA용Lipofectamine 2000전염BMSCs、분위전염조、공백대조조화음성대조조，응용실시정량PCR、Western blotting、격광공취초현미관찰등방법검측siRNA대예분화BMSCs적β2 M mRNA화단백적표체정황，갑분알람화Ⅱ형효원면역형광검측예분화BMSC적단백취당다취체화Ⅱ형효원분비정황。결과：실시정량PCR、Western blotting 화격광공취초현미관찰결과현시siRNA성공억제β2 M mRNA화단백적표체，갑분알람화Ⅱ형효원면역형광결과현시siRNA불영향예분화BMSCs단백취당다취체화Ⅱ형효원단백적분비。결론： RNAi 간우β2 M 가강저BMSCs 중β2 M mRNA화단백적표체，단불영향예분화BMSCs적연골세포특성。
[ABSTRACT]AIM:TostudytheeffectofsmallinterferingRNA(siRNA)ontheexpressionofbeta2-microglo-bulin (β2M) in pre-differentiated bone marrow mesenchymal stem cells (BMSCs).METHODS: The β2M siRNA was transfected into the pre-differentiated BMSCs with Lipofectamine 2000.BMSCs were divided into transfection group , blank control group and negative control group .The expression of β2 M at mRNA and protein levels was determined by real-time qPCR, Western blotting and laser confocal microscopy .The productions of aggrecan and type II collagen in pre-differentia-ted BMSCs were determined by toluidine blue staining and type Ⅱcollagen immunofluorescence .RESULTS:The results of real-time qPCR, Western blotting and laser confocal microscopy showed that siRNA successfully inhibited the expression ofβ2 M at mRNA and protein levels in the pre-differentiated BMSCs .The results of toluidine blue and type Ⅱcollagen im-munofluorescence staining showed that siRNA does not affect the productions of aggrecan and type Ⅱ collagen in the pre-differentiated BMSCs .CONCLUSION:siRNA targeting β2 M reduces the expression of β2 M in the pre-differentiated BM-SCs and does not affect the chondrocyte characteristics of pre -differentiated BMSCs .