CAJ | 학술논문

目的：检测髓样细胞白血病-1基因（myeoid cell leukemin-1，Mcl-1）在胃癌细胞SGC7901及MGC-803中的表达，并筛选出最有效的siRNA序列。方法：RT-PCR检测胃癌细胞中Mcl-1基因的表达；设计针对Mcl-1的4对干扰序列（Mcl-1siRNA 1～4）及1对阴性对照（Mcl-1siRNA NC）。瞬时转染胃癌细胞，荧光定量PCR和Western blot分别检测转染24、48、72 h后细胞中Mcl-1含量。结果：2种胃癌细胞均存在Mcl-1基因的表达。综合分析，SGC7901及MGC-803细胞转染48 h后达到最佳抑制效果；以Mcl-1siRNA1为最好， Mcl-1 mRNA的抑制率分别为73．8％、67．6％；蛋白抑制率分别为79．3％和96．1％。结论：本研究成功设计并筛选出能够同时有效抑制2种胃癌细胞Mcl-1表达的siRNA1序列。
목적：검측수양세포백혈병-1기인（myeoid cell leukemin-1，Mcl-1）재위암세포SGC7901급MGC-803중적표체，병사선출최유효적siRNA서렬。방법：RT-PCR검측위암세포중Mcl-1기인적표체；설계침대Mcl-1적4대간우서렬（Mcl-1siRNA 1～4）급1대음성대조（Mcl-1siRNA NC）。순시전염위암세포，형광정량PCR화Western blot분별검측전염24、48、72 h후세포중Mcl-1함량。결과：2충위암세포균존재Mcl-1기인적표체。종합분석，SGC7901급MGC-803세포전염48 h후체도최가억제효과；이Mcl-1siRNA1위최호， Mcl-1 mRNA적억제솔분별위73．8％、67．6％；단백억제솔분별위79．3％화96．1％。결론：본연구성공설계병사선출능구동시유효억제2충위암세포Mcl-1표체적siRNA1서렬。
Objective To detect the expression of Mcl-1 gene in gastric cancer cell lines SGC7901 and MGC-803, and to screen the most effective Mcl-1-targeted siRNA sequence. Methods Mcl-1 expression was evaluated in human gastric cancer cell lines SGC7901 and MGC-803 by RT-PCR. Four segments of siRNAs (siRNA1, siRNA2, siRNA3 and siRNA4) targeting Mcl-1 mRNA and a no-sense control segment were designed by bioinformatic technology . Mcl-1 specific siRNAs were transfected transiently into SGC7901 and MGC-803 cells by using lipofectamine 2000 . After transfected 24 , 48 and 72 h , quantitative real-time PCR was applied to detect the mRNA expression of Mcl-1 and western-blot analysis was applied to detect the protein expression. Results Mcl-1 gene was expressed in both SGC7901 and MGC-803 cells. Overall, siRNA1 exhibited the best inhibitory effect after being transfected for 48h. The inhibition rate of mRNA level in SGC7901 group and MGC-803 group was 73.8%and 67.6%, and the inhibition rate of protein level in SGC7901 group and MGC-803 group was 79.3%and 96.1%. Conclusion Mcl-1 specific siRNA sequences were successfully designed, and siRNA1 was selected as the most effective sequence, which can simultanandeously inhibit the expression of Mcl-1 in GC7901 and MGC-803 cells.