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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 investigate whether estrogen regulates the expression of glyoxalase Ⅰ (Glo Ⅰ) in KLE cells of endometrial carcinoma by AKT pathway. Methods: The expression of GloⅠ mRNA and protein in endometrial carcinoma KLE cells was detected by RT-PCR and Western blotting after estrogen (17-β estradiol) treatment or AKT pathway inhibitor (LY294002) treatment. The experiment was divided into 4 groups: Con group (control group); LY294002 group (LY294002 treatment group); E2 group (17-β estradiol treatment group); E2 + LY294002 group Alcohol treatment group). The results showed that the relative expression of GloⅠmRNA in KLE cells treated with different concentrations of 17-β estradiol (Con, 10-11,10-10,10-9M) for 24 hours were 1, 1.5 ± 0.04, 1.82 ± 0.03 and 1.81 ± 0.04, the most significant at 10-10 M (P <0.05). After 48 hours of treatment with 10-10 M 17-beta estradiol in KLE cells, the relative expression level of Glo I protein was 1.79 ± 0.02, which was higher than that of Con group. After LY294002 inhibited AKT pathway, the relative expression of GloⅠmRNA in KLE cells was 0.69 ± 0.03, and the relative expression of protein was 0.16 ± 0.02, which were all lower than those in Con group. The relative expression levels of GloⅠmRNA and protein in E2 + LY294002 group were 1.02 ± 0.04 and 1.01 ± 0.03, respectively, which were all lower than those in E2 group (E2 = 1.34 ± 0.03,1.79 ± 0.02, respectively). The relative expression levels of GloⅠ mRNA and protein in LY294002 group were 0.69 ± 0.03,0.16 ± 0.02, which were all lower than those in Con group. Conclusion: Estrogen can up-regulate the expression of GloⅠmRNA and protein in KLE cells of endometrial carcinoma. Inhibition of AKT pathway downregulates Glo Ⅰ mRNA and protein expression in KLE cells of endometrial cancer. AKT pathway in the estrogen regulation of endometrial cancer KLE cells Glo Ⅰ expression had no significant effect.