中国糖尿病杂志
中國糖尿病雜誌
중국당뇨병잡지
CHINESE JOURNAL OF DIABETES
2010年
12期
934-937
,共4页
沙建平%薛耀明%陈炫%罗祥蓉%关美萍%曾展军%何飞英%王玲%毛睿睿
沙建平%薛耀明%陳炫%囉祥蓉%關美萍%曾展軍%何飛英%王玲%毛睿睿
사건평%설요명%진현%라상용%관미평%증전군%하비영%왕령%모예예
RNAi%胰岛新生相关蛋白%细胞增殖
RNAi%胰島新生相關蛋白%細胞增殖
RNAi%이도신생상관단백%세포증식
RNA interference%Islet neogenesis associated protein （INGAP）%Cell proliferation
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 mR-NA和蛋白表达的变化,用噻唑蓝(MTT)法来检测细胞的增殖.结果 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.
目的 探討靶嚮胰島新生相關蛋白(INGAP)基因RNAi對胰島細胞增殖的抑製效應.方法 設計閤成siRNA,通過脂質體轉入胰島細胞株INS-1中,研究其對靶基因的抑製效應,採用逆轉錄-聚閤酶鏈反應(RT-PCR)、流式細胞儀、Western-blot法分彆檢測轉染後的INS-1細胞中INGAP mR-NA和蛋白錶達的變化,用噻唑藍(MTT)法來檢測細胞的增殖.結果 siRNA6序列對細胞增殖有明顯的抑製效應,INGAP mRNA及蛋白錶達明顯降低,INS-1細胞增殖受到抑製(P＜0.05).結論 榦擾INGAP基因的錶達能有效抑製胰島細胞的增殖,INGAP有可能成為治療β細胞嚴重減少或損害的重要新靶點.
목적 탐토파향이도신생상관단백(INGAP)기인RNAi대이도세포증식적억제효응.방법 설계합성siRNA,통과지질체전입이도세포주INS-1중,연구기대파기인적억제효응,채용역전록-취합매련반응(RT-PCR)、류식세포의、Western-blot법분별검측전염후적INS-1세포중INGAP mR-NA화단백표체적변화,용새서람(MTT)법래검측세포적증식.결과 siRNA6서렬대세포증식유명현적억제효응,INGAP mRNA급단백표체명현강저,INS-1세포증식수도억제(P＜0.05).결론 간우INGAP기인적표체능유효억제이도세포적증식,INGAP유가능성위치료β세포엄중감소혹손해적중요신파점.
Objective To investigate the effects of small interfering RNA (siRNA) on the expression of islet neogenesis associated protein (INGAP) and on cell propagation of INS-1 cells. Methods Various siRNAs against the INGAP gene were designed and transfected into islet cells lines INS-1, respectively . The expression levels of INGAP mRNA and protein were detected by RT-PCR, flow cytometry and Western blotting, respectively. The proliferation of INS-1 cells was detected by MTT. Results Compared with irrelevant siRNA6 control group, empty vector control group and un-transfected group,the expression levels of INGAP mRNA and protein were obviously reduced in expermiental groups (all P＜005). The cell proliferation speed significantly increased in transfection group (P<005). Conclusions siRNA against INGAP could effectively down-regulates INGAP expression in INS-1cell line, and it can inhibit cell proliferation.
