AKT途径在雌激素调控子宫内膜癌KLE细胞中乙二醛酶Ⅰ表达的作用

A Possible Role of AKT Signal Pathway for Estrogen Regulates Glyoxalase Ⅰ Gene Expression in KLE Endometrial Cancer Cells

目的:探讨雌激素是否通过AKT途径调控子宫内膜癌KLE细胞中乙二醛酶Ⅰ(GlyoxalaseⅠ,GloⅠ)的表达。方法:采用RT-PCR和Western blotting检测雌激素(17-β雌二醇)处理或AKT途径抑制剂(LY294002)处理子宫内膜癌KLE细胞后GloⅠ的mRNA或蛋白的表达情况。实验分4组:Con组(对照组);LY294002组(LY294002处理组);E2组(17-β雌二醇处理组);E2+LY294002组(LY294002预处理1小时后再17-β雌二醇处理组)。结果:1、经不同浓度的17-β雌二醇(Con、10-11、10-10、10-9M)作用于KLE细胞24小时后,GloⅠmRNA的相对表达量分别为1,1.58±0.04,1.82±0.03,1.81±0.04,以10-10 M的浓度时最为显著(P<0.05)。以10-10 M的17-β雌二醇作用于KLE细胞48小时后,GloⅠ蛋白的相对表达量为1.79±0.02,高于Con组。2、以LY294002抑制AKT途径后,KLE细胞中GloⅠmRNA的相对表达量为0.69±0.03,蛋白的相对表达量为0.16±0.02,均低于Con组。3、E2+LY294002组GloⅠmRNA和蛋白的相对表达量分别为1.02±0.04、1.01±0.03,均低于E2组中GloⅠmRNA和蛋白的相对表达量,E2组分别为1.34±0.03、1.79±0.02。LY294002组GloⅠmRNA和蛋白的相对表达量分别为0.69±0.03,0.16±0.02,均低于Con组GloⅠmRNA和蛋白的相对表达量。结论:雌激素可上调子宫内膜癌KLE细胞中GloⅠmRNA和蛋白的表达。抑制AKT途径可下调子宫内膜癌KLE细胞中GloⅠmRNA和蛋白的表达。AKT途径在雌激素调控子宫内膜癌KLE细胞中GloⅠ的表达无明显作用。

Objective： To study the possible role of AKT signal pathway for estrogen regulates Glyoxalase Ⅰ Gene expression in KLE endometrial cancer cells. Methods： RT-PCR and western blot were used to determine the effect of estrogen or ly294002 on GloⅠ expression in mRNA and protein levels respectively. KLE cells were divided into 4 groups： Con group （the control group）; LY294002 group（which were treated by LY294002）;E2 group（which were treated by E2）;E2 ＋LY294002 group（which were treated by LY294002 for 1 h,then with E2）. Results： 1 .The expression of GloⅠ mRNA levels in KLE cells, which were treated by E2 （Con, 10^-11, 10^-10, 10^-9 M） for 24h, were increased. The results were：l, 1.58± 0.04, 1.82±0.03,1.81±0.04（P〈0.05 ）. The protein levels were also increased, the results were 1.79±0.02 （P〈0.05）. 2 After treated by LY294002, the expression ofGloⅠ mRNA and protein levels were decreased. The results were 0.69±0.03 and 0.16±0.02 （P〈0.05）.3.The expression of GloⅠ mRNA and protein levels of E2 ＋LY294002 group were lower than E2 group. The difference between the group： LY294002 and Con were the same. Conclusions： E2 up-regulated the expression of GloⅠ . Inhibition of AKT signal pathway by LY294002 reduced the expression of GloⅠ . AKT signal pathway is not the main step for estrogen regulates Glyoxalase Ⅰ Gene expression in KLE endometrial cancer cells.