CAJ | 학술논문

目的探讨靶向胰岛新生相关蛋白(INGAP)基因RNAi对胰岛细胞增殖的抑制效应.方法设计合成siRNA,通过脂质体转入胰岛细胞株INS-1中,研究其对靶基因的抑制效应,采用逆转录.聚合酶链反应(RT-PCR)、流式细胞仪、Western-blot法分别检测转染后的INS-1细胞中INGAP mRNA和蛋白表达的变化,用噻唑蓝(MTT)法来检测细胞的增殖.结果结果显示siRNA6序列对细胞增殖有明显的抑制效应,INGAP mRNA及蛋白表达明显降低,INS-1细胞增殖受到抑制,P<0.05.结论干扰INGAP基因的表达能有效抑制胰岛细胞的增殖,INGAP有可能成为治疗β细胞严重减少或损害的重要新靶点.
목적 탐토파향이도신생상관단백(INGAP)기인RNAi대이도세포증식적억제효응.방법 설계합성siRNA,통과지질체전입이도세포주INS-1중,연구기대파기인적억제효응,채용역전록.취합매련반응(RT-PCR)、류식세포의、Western-blot법분별검측전염후적INS-1세포중INGAP mRNA화단백표체적변화,용새서람(MTT)법래검측세포적증식.결과 결과현시siRNA6서렬대세포증식유명현적억제효응,INGAP mRNA급단백표체명현강저,INS-1세포증식수도억제,P<0.05.결론 간우INGAP기인적표체능유효억제이도세포적증식,INGAP유가능성위치료β세포엄중감소혹손해적중요신파점.
Objective To investigate the effect of small interfering RNA (siRNA)-mediated islet neogenesis associated protein (INGAP) gene silencing on the proliferation of islet cells. Methods Different siRNAs targeting INGAP gene were designed and transfected into INS-1 islet cells, and the expression levels of INGAP mRNA and protein following the transfection were detected using RT-PCR, flow cytometry and Western blotting. The proliferation of the transfected INS-1 cells was evaluated using MTT assay. Results Compared with those in the irrelevant siRNA, empty vector control, and un-transfected groups, the expression levels of INGAP mRNA and protein in the cells transfected with siRNA6 were reduced significantly. The cell proliferation rate significantly increased after transfection with siRNA6 (P<0.05). Conclusion siRNA targeting INGAP can effectively down-regulate INGAP expression and inhibit the proliferation of INS-1 cells.