目的:研究星形细胞上调基因1(astrocyte elevated gene-1,AEG-1)在人宫颈鳞癌细胞和组织中的表达,探讨AEG-I表达下调对宫颈癌SiHa细胞细胞周期和侵袭能力的影响,并分析其可能的分子机制.方法:采用Western blotting检测正常宫颈组织、宫颈鳞癌组织、HeLa、SiHa和CaSki细胞中AEG-1蛋白的表达.将对照siR-NA和AEG-1 siRNA分别转染SiHa细胞,利用Western blotting检测SiHa细胞中AEG-1蛋白的表达,采用流式细胞术检测细胞周期分布的变化,采用Boyden小室检测细胞侵袭能力的变化,最后采用Western blotting检测cyclin D1、细胞周期素依赖性激酶2(CDK2)和基质金属蛋白酶9(MMP-9)蛋白表达的变化.结果:宫颈鳞癌组织中AEG-1蛋白表达显著高于正常宫颈组织(P＜0.05),同时3株宫颈癌细胞中AEG-1蛋白表达均显著高于正常宫颈组织,其中SiHa细胞中AEG-1蛋白表达最高(P＜0.05).此外,AEG-1 siRNA能显著下调SiHa细胞中AEG-1蛋白的表达(P＜0.05),其表达下调能明显促使SiHa细胞在G0/G1期的比例增加和降低其侵袭能力.Western blotting结果表明,AEG-1 siRNA组中cyclin D1、CDK2和MMP-9蛋白的表达均显著低于未处理组和对照siRNA组(P＜0.05).结论:AEG-1在宫颈癌中的高表达可能与宫颈癌的发生发展密切相关,其表达下调介导的细胞周期静止和侵袭能力降低可能与cyclin D1、CDK2和MMP-9蛋白表达下调密切相关.%AIM:To investigate the effects of down-regulation of astrocyte elevated gene-1 (AEG-1) expression on cell cycle and invasion of human cervical carcinoma SiHa cells.METHODS:The protein expression of AEG-1 was detected by Western blotting in normal cervical tissues,cervical squamous cell carcinoma tissues,HeLa cells,SiHa cells and CaSki cells.Control siRNA or AEG-1 siRNA was transfected into SiHa cells,and the protein expression of AEG-1 in SiHa cells was detected by Western blotting.The changes of cell cycle distribution and cell invasion were determined by flow eytometry and Boyden chamber,respectively.The protein levels of cyclin D1,cyclin-dependent kinase 2 (CDK2) and matrix metalloproteinase-9 (MMP-9) were analyzed by Western blotting.RESULTS:The protein expression of AEG-1 in cervical squamous cell carcinoma tissues was significantly higher than that in normal cervical tissues (P ＜ 0.05).Meanwhile,the protein expression of AEG-1 in the 3 cervical carcinoma cell lines was obviously higher than that in normal cervical tissues,in which SiHa cells displayed the highest AEG-1 protein level (P ＜ 0.05).In addition,AEG-1 siRNA effectively down-regulated the protein expression of AEG-1 in SiHa cells,which led to increase the percentage at G0/G1 phase and reduced the invasion of SiHa cells.Furthermore,the protein levels of cyclin D1,CDK2 and MMP-9 in AEG-1 siRNA=group were markedly lower than those in non-treatment group and control siRNA group (P ＜ 0.05).CONCLUSION:Over-expression of AEG-1 may be closely associated with the occurrence and development of cervical carcinoma,and the AEG-1 down-regulation-mediated cell cycle arrest and attenuation of invasion may be tightly related to the down-regulations of cyclin D1,CDK2 and MMP-9 at protein levels.
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