中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
1期
22-27
,共6页
张彩凤%夏永华%郑庆芬%李贞娟%郭晓鹤%周慧聪%张利利%董良鹏%韩宇
張綵鳳%夏永華%鄭慶芬%李貞娟%郭曉鶴%週慧聰%張利利%董良鵬%韓宇
장채봉%하영화%정경분%리정연%곽효학%주혜총%장리리%동량붕%한우
胃肿瘤%星形细胞上调基因1%RNA,小分子干扰%细胞增殖
胃腫瘤%星形細胞上調基因1%RNA,小分子榦擾%細胞增殖
위종류%성형세포상조기인1%RNA,소분자간우%세포증식
Stomach neoplasms%Astrocyte elevated gene-1%RNA,small interfering%Cell proliferation
目的 探讨星形细胞上调基因1(AEG-1)表达水平对胃癌细胞增殖和细胞周期的影响,研究其可能的分子机制.方法 以siRNA和AEG-1 siRNA转染胃癌SGC-7901细胞后48 h,将细胞分为未转染组、siRNA对照组和AEG-1 siRNA转染组.应用实时荧光定量PCR和Western blot检测转染后SGC-7901细胞中AEG-1 mRNA和蛋白的表达,应用CCK-8检测细胞增殖能力,应用流式细胞术检测转染前后细胞周期的分布,应用Western blot检测细胞增殖和细胞周期相关蛋白表达的变化.结果 与未转染组和siRNA对照组比较,AEG-1 siRNA转染组胃癌细胞中AEG-1蛋白的表达水平下降(P＜0.05).与未转染组和siRNA对照组比较,AEG-1 siRNA转染组AEG-1 mRNA表达水平下降(P＜0.05).与未转染组和siRNA对照组比较,AEG-1 siRNA转染组转染AEG-1后的24h及其后续的各个时间点,SGC-7901细胞的增殖均明显受到抑制(均P＜0.05).AEG-1 siRNA转染组中G0/G1期细胞数比例[(61.26±1.25)％]显著高于未转染组[(46.17±1.91)％]和siRNA对照组[(46.46±1.96)％],差异均有统计学意义(均P ＜0.05).AEG-1表达水平下降可使cdk2和cyclinD1表达水平下降以及p21表达水平升高.结论 AEG-1表达水平下降抑制胃癌细胞的增殖和改变细胞周期分布,可能与cdk2、cyclin D1和p21蛋白的表达密切相关.
目的 探討星形細胞上調基因1(AEG-1)錶達水平對胃癌細胞增殖和細胞週期的影響,研究其可能的分子機製.方法 以siRNA和AEG-1 siRNA轉染胃癌SGC-7901細胞後48 h,將細胞分為未轉染組、siRNA對照組和AEG-1 siRNA轉染組.應用實時熒光定量PCR和Western blot檢測轉染後SGC-7901細胞中AEG-1 mRNA和蛋白的錶達,應用CCK-8檢測細胞增殖能力,應用流式細胞術檢測轉染前後細胞週期的分佈,應用Western blot檢測細胞增殖和細胞週期相關蛋白錶達的變化.結果 與未轉染組和siRNA對照組比較,AEG-1 siRNA轉染組胃癌細胞中AEG-1蛋白的錶達水平下降(P＜0.05).與未轉染組和siRNA對照組比較,AEG-1 siRNA轉染組AEG-1 mRNA錶達水平下降(P＜0.05).與未轉染組和siRNA對照組比較,AEG-1 siRNA轉染組轉染AEG-1後的24h及其後續的各箇時間點,SGC-7901細胞的增殖均明顯受到抑製(均P＜0.05).AEG-1 siRNA轉染組中G0/G1期細胞數比例[(61.26±1.25)％]顯著高于未轉染組[(46.17±1.91)％]和siRNA對照組[(46.46±1.96)％],差異均有統計學意義(均P ＜0.05).AEG-1錶達水平下降可使cdk2和cyclinD1錶達水平下降以及p21錶達水平升高.結論 AEG-1錶達水平下降抑製胃癌細胞的增殖和改變細胞週期分佈,可能與cdk2、cyclin D1和p21蛋白的錶達密切相關.
목적 탐토성형세포상조기인1(AEG-1)표체수평대위암세포증식화세포주기적영향,연구기가능적분자궤제.방법 이siRNA화AEG-1 siRNA전염위암SGC-7901세포후48 h,장세포분위미전염조、siRNA대조조화AEG-1 siRNA전염조.응용실시형광정량PCR화Western blot검측전염후SGC-7901세포중AEG-1 mRNA화단백적표체,응용CCK-8검측세포증식능력,응용류식세포술검측전염전후세포주기적분포,응용Western blot검측세포증식화세포주기상관단백표체적변화.결과 여미전염조화siRNA대조조비교,AEG-1 siRNA전염조위암세포중AEG-1단백적표체수평하강(P＜0.05).여미전염조화siRNA대조조비교,AEG-1 siRNA전염조AEG-1 mRNA표체수평하강(P＜0.05).여미전염조화siRNA대조조비교,AEG-1 siRNA전염조전염AEG-1후적24h급기후속적각개시간점,SGC-7901세포적증식균명현수도억제(균P＜0.05).AEG-1 siRNA전염조중G0/G1기세포수비례[(61.26±1.25)％]현저고우미전염조[(46.17±1.91)％]화siRNA대조조[(46.46±1.96)％],차이균유통계학의의(균P ＜0.05).AEG-1표체수평하강가사cdk2화cyclinD1표체수평하강이급p21표체수평승고.결론 AEG-1표체수평하강억제위암세포적증식화개변세포주기분포,가능여cdk2、cyclin D1화p21단백적표체밀절상관.
Objective To explore the effect of down-regulation of astrocyte elevated gene-1 (AEG-1 expression on cell proliferation and cell cycle of gastric carcinoma cells,and its possible molecular mechanism.Methods Control siRNA and AEG-1 siRNA were transfected into gastric carcinoma SGC-7901 cells.48 h after transfection,the cells were divided into 3 groups including untransfected,siRNA control and AEG-1 siRNA transfection groups.Expressions of AEG-1 mRNA and protein in the 3 group cells were detected by real-time quantitative PCR and Western blot.The changes of cell proliferation were examined using CCK-8 kit,and the cell cycle distribution was detected by flow cytometry.Finally,expressions of cell proliferation and cell cycle related proteins were detected by Western blot.Results Real-time quantitative PCR and Western blot demonstrated that compared with the untransfected and siRNA control groups,expressions of AEG-1 mRNA and protein were significantly down-regulated in the AEG-1 siRNA transfection group (P ＜ 0.05),but there was no significant difference between the untransfected and siRNA control groups (P ＞ 0.05).Furthermore,in vivo experiment confirmed a significant down-regulation of AEG-1 protein in the AEG-1 siRNA transfection group (P ＜ 0.05).In addition,AEG-1 siRNA obviously inhibited the proliferation of SGC-7901 cells at different time points after transfection with AEG-1 siRNA.The percentage of cells in G0/G1 phase in the AEG-1 siRNA transfection group [(61.26 ± 1.25) ％] was significantly higher than those in the untransfected group [(46.17 ± 1.91) ％] and siRNA control group [(46.46 ± 1.96) ％],and there was a significant difference between them (all P ＜ 0.001).Furthermore,the result of Western blotting revealed that down-regulation of AEG-1 expression evoked the down-regulation of cdk2 and cyclin D1 expressions and elevation of p21 expression in vitro and in vivo.Conclusions The inhibition of cell proliferation and cell cycle arrest mediated by down-regulation of AEG-1 expression may be closely associated with the changes of expression of cell cycle related proteins including cdk2,cyclin D1 and p21.
