CAJ | 학술논문

目的探讨胰岛素样生长因子受体(IGF-1R)基因干扰对人胃癌细胞株MKN28增殖的影响及其机制.方法构建针对IGF-1R的小干扰RNA(siRNA-IGF-1R),采用Western blot检测转染后MKN28细胞IGF-1R、磷酸肌醇3激酶(PI3K)、蛋白激酶B(Akt)、p21、整合酶相互作用分子1(INI1)的表达变化.流式细胞仪检测各组细胞生长周期.噻唑蓝(MTT)法检测MKN28细胞株的增殖.结果与对照组比较,siRNA组IGF-1R、PI3K及Akt的表达水平均显著降低,同时p21及INI1表达水平升高.流式细胞仪检测细胞周期和MTT实验结果显示,G0/G1期细胞比例显著升高,S期细胞比例显著降低,细胞增殖能力被显著抑制,第2～4天siRNA-IGF-1R对细胞增殖的抑制率分别为31.12％、36.22％和42.03％,差异有统计学意义(P＜0.05).结论抑制IGF-1R基因表达可下调PI3K、Akt表达,上调p21、INI1表达,从而有效抑制MKN28细胞的增殖能力.
목적 탐토이도소양생장인자수체(IGF-1R)기인간우대인위암세포주MKN28증식적영향급기궤제.방법 구건침대IGF-1R적소간우RNA(siRNA-IGF-1R),채용Western blot검측전염후MKN28세포IGF-1R、린산기순3격매(PI3K)、단백격매B(Akt)、p21、정합매상호작용분자1(INI1)적표체변화.류식세포의검측각조세포생장주기.새서람(MTT)법검측MKN28세포주적증식.결과 여대조조비교,siRNA조IGF-1R、PI3K급Akt적표체수평균현저강저,동시p21급INI1표체수평승고.류식세포의검측세포주기화MTT실험결과현시,G0/G1기세포비례현저승고,S기세포비례현저강저,세포증식능력피현저억제,제2～4천siRNA-IGF-1R대세포증식적억제솔분별위31.12％、36.22％화42.03％,차이유통계학의의(P＜0.05).결론 억제IGF-1R기인표체가하조PI3K、Akt표체,상조p21、INI1표체,종이유효억제MKN28세포적증식능력.
Objective To study the effects of insulin-like growth factor (IGF)-1R interference on proliferation of MKN28 gastric cancer cells.Methods Small interfering RNA (siRNA) targeting IGF-1 R mRNA was obtained by in vitro transcription and transfected into cells with lipofectin.The survival of the transfected cells was detected by using methyl thiazol tetrazolium (MTT) assay.The IGF-1R,phosphatidylinositol 3 kinase (PI3K),protein kinase B (Akt),p21 and integrase interacting molecule 1 (INI1) expression levels were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.Cell cycle of siRNA group was determined by flow cytometry (FCM).Results Inhibition of IGF-1R effectively inhibited the proliferation of MKN28 cells.The expression levels of IGF-1 R,PI3K and Akt mRNA and proteins were significantly inhibited,while those of p21 and INI1 were up-regulated in MKN28 cells in siRNA group as compared with control group.The percentage of cells in G0/G1 phase was significantly higher in siRNA group than in control group,2-4 d the cell proliferation rate of siRNA-IGF-1R were 31.12％,36.22％ and 42.03％,the difference has statistical significance (P ＜ 0.05).Conclusion IGF-I R gene interference could inhibit proliferation of MKN28 cells by regulating PI3K,Akt,p21 and INI1 expression.