姜黄素对子宫内膜癌孕激素抵抗的影响

doi:10.6040/j.issn.1671-7554.0.2020.1753

摘要/Abstract

摘要： 目的观察不同浓度姜黄素对子宫内膜癌孕激素耐药细胞的影响,探讨相关信号通路在影响子宫内膜癌孕激素耐药中的作用。方法采用ETCM、GEPIA数据库分析姜黄素的靶点基因,进而筛选出与子宫内膜癌及耐药相关的基因;测定已建立的子宫内膜癌Ishikawa细胞的孕激素耐药模型(IshikawaPR细胞系)的耐药性,将耐药细胞按不同浓度处理分为姜黄素组(给予0、5、10、20、40、60、80 μmoL姜黄素)、孕激素组(给予0、5、10、20、40、60、90 μmoL孕激素)和姜黄素+孕激素组(同时给予相同浓度的姜黄素和孕激素),采用噻唑蓝溴化四唑(MTT)检测各组细胞的生长;采用5-乙炔基-2'-脱氧尿苷(EdU)、小室穿透实验(Transwell)、线粒体膜电位检测分析姜黄素对子宫内膜癌耐药细胞的影响;数据库分析姜黄素靶向作用的蛋白,采用Western blotting检测不同浓度姜黄素刺激后耐药细胞内蛋白水平的变化,并结合数据库分析潜在的信号通路。结果数据库分析结果显示,姜黄素靶点基因与PGR、ESR等肿瘤耐药基因密切相关;MTT结果显示,姜黄素能够抑制耐药细胞生长,导致细胞存活率下降(F=907.8, P<0.001),并且姜黄素和孕激素之间存在交互关系(F=51.34,P=0.004),当孕激素给药浓度分别为10 μmoL(t=16.13,P=0.026)、20 μmoL(t=33.23,P=0.006)、40 μmoL(t=35.94,P =0.005)时,联合姜黄素可较孕激素单药处理使耐药细胞抑制效率更强。姜黄素(0、10、20 μmoL)处理可抑制耐药细胞增殖(F=205.8, P<0.001)、侵袭(F=13.9,P=0.006),促进细胞凋亡(F=23.3,P=0.002);并且耐药细胞中CyclinD1、CASP3、MMP2、磷酸化的核内转录因子蛋白水平下降,孕激素受体表达升高。结论姜黄素可抑制子宫内膜癌孕激素抵抗,可能是通过介导转录因子NF-κB 失活实现。

关键词: 姜黄素, 子宫内膜癌, 孕激素, 耐药, 核转录因子NF-κB

Abstract: Objective To explore the effects of different concentrations of curcumin on progesterone resistant cells in endometrial carcinoma and to investigate the role of related signaling pathways influencing progesterone resistance. Methods The target genes of curcumin were analyzed in the ETCM and GEPIA databases and the interaction genes related to tumor-drug resistance were screened out. The resistance of an established progesterone-resistance model of Ishikawa cells in endometrial carcinoma(IshikawaPR cell line)was detected. Resistant cells were divided into curcumin(0, 5, 10, 20, 40, 60, 80 μmoL)groups, progesterone(0, 5, 10, 20, 40, 60, 90 μmoL)groups and curcumin+progesterone(the same concentration)groups, according to different treatments. The proliferation of cells in each group was determined with MTT. The effects of curcumin on the resistant cells were assessed with EDU, Transwell and mitochondrial membrane potential. The changes of intracellular protein levels after curcumin stimulation at different concentrations were determined with Western blotting. The potential signaling pathways were analyzed in the databases. Results Analyses of the databases indicated that curcumin target genes were closely related to tumor-drug resistance genes such as PGR and ESR. MTT results showed that curcumin inhibited the growth of drug-resistant cells, leading to a decrease in cell survival rate(F=907.8, P<0.001), and there was an interactive relationship between curcumin and progesterone(F=51.34, P=0.004). When the concentrations of progesterone were 10 μmoL(t=16.13, P=0.026), 20 μmoL(t=33.23, P=0.006), and 40μmoL(t=35.94, P=0.005), combination with curcumin inhibited drug-resistant cells more effectively than the use of progesterone alone. Curcumin treatment(0, 10, 20 μmoL)inhibited the proliferation(F=205.8, P<0.001), invasion(F=13.9, P=0.006)and promoted apoptosis(F=23.3, P=0.002)of drug-resistant cells. Western blotting showed that after curcumin treatment, the levels of CyclinD1, CASP3, MMP2 and p-NFκB decreased while the level of PR increased in resistant cells. Conclusion Curcumin inhibits progesterone resistance in endometrial carcinoma, possibly by mediating inactivation of transcription factor NFκB.

Key words: Curcumin, Endometrial carcinoma, Progesterone, Resistance, Nuclear transcription factor-κB

中图分类号:

R711

丁菲,姜洁. 姜黄素对子宫内膜癌孕激素抵抗的影响[J]. 山东大学学报 (医学版), 2021, 59(4): 35-41.

DING Fei, JIANG Jie. Effects of curcumin on progesterone resistance in endometrial carcinoma[J]. Journal of Shandong University (Health Sciences), 2021, 59(4): 35-41.

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