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目的:探讨PGE2对子宫内膜癌细胞生长的促进作用及分子机制。方法:免疫组织化学(EnVision)法检测正常子宫内膜与子宫内膜癌组织中β-catenin的表达;用PGE2、EP2受体激动剂Butaprost、EP2受体拮抗剂AH6809处理子宫内膜癌Ishikawa细胞株,以WST-8细胞增殖实验检测各实验组细胞的增殖率;用Western blot方法分别检测β-catenin、p-β-catenin、GSK-3β、p-GSK-3β在未处理组、PGE2处理组、EP2受体激动剂处理组和EP2受体拮抗剂处理组中的表达;免疫荧光技术检测β-catenin在未处理组和PGE2处理组中表达定位的情况。结果:免疫组织化学法结果显示β-catenin在正常子宫内膜组织中仅存在膜表达,而子宫内膜癌组织中为胞浆和胞核的表达。WST-8实验结果显示:经PGE2、Butaprost和AH6809处理后,细胞的增殖率相对于对照组分别为183.9%、151.9%、72.1%。Western blot实验结果显示:PGE2处理后,β-catenin蛋白表达上升至235.7%而p-β-catenin水平下降至76.8%,p-GSK-3β水平升高至204.3%;EP2受体激动剂+PGE2处理组显示β-catenin蛋白表达上升至279.7%而p-β-catenin水平下降至33.6%,p-GSK-3β水平升高至236.7%;EP2受体拮抗剂+PGE2处理组显示β-catenin蛋白表达上升至185.2%而p-β-catenin水平下降至73.9%,p-GSK-3β水平升高至116.2%。免疫荧光技术探测结果表明:PGE2处理子宫内膜癌细胞后,β-catenin蛋白在癌细胞胞浆中的表达信号明显强于未处理组,并出现明显核内表达信号。结论:PGE2能促进子宫内膜癌细胞的增殖,并伴有β-catenin蛋白的过表达和入核,其机制可能涉及到EP2受体激活和Wnt/β-catenin信号转导通路。
Objective: To investigate the effect of PGE2 on the growth of endometrial carcinoma cells and its molecular mechanism. Methods: EnVision method was used to detect the expression of β-catenin in normal endometrium and endometrial carcinoma tissues. Ishikawa cells were treated with PGE2, EP2 receptor agonist Butaprost and EP2 receptor antagonist AH6809 The proliferation of cells in each experimental group was detected by WST-8 cell proliferation assay. The expression of β-catenin, GSK-3β and p-GSK-3β in untreated group and PGE2 EP2 receptor agonist group and EP2 receptor antagonist group. The expression of β-catenin in untreated and PGE2-treated groups was detected by immunofluorescence staining. Results: The results of immunohistochemistry showed that there was only membrane expression of β-catenin in normal endometrial tissue, while the expression of cytoplasm and nucleus in endometrial carcinoma tissue. The results of WST-8 showed that the proliferation rate of cells treated with PGE2, Butaprost and AH6809 were 183.9%, 151.9% and 72.1% respectively. The results of Western blot showed that after PGE2 treatment, the protein expression of β-catenin increased to 235.7%, the level of p-β-catenin decreased to 76.8% and the level of p-GSK-3β increased to 204.3% The expression of β-catenin protein increased to 279.7% and the level of p-β-catenin decreased to 33.6% and the level of p-GSK-3β increased to 236.7% The expression increased to 185.2%, the level of p-β-catenin decreased to 73.9%, and the level of p-GSK-3β increased to 116.2%. The results of immunofluorescence showed that the expression of β-catenin protein in cytoplasm of cancer cells was stronger than that of untreated cells after PGE2 treatment, and the signal of nuclear expression was obvious. CONCLUSION: PGE2 can promote the proliferation of endometrial cancer cells accompanied by overexpression and nuclear import of β-catenin protein, which may involve the activation of EP2 receptor and Wnt / β-catenin signal transduction pathway.