山东大学学报 (医学版) ›› 2022, Vol. 60 ›› Issue (5): 74-80.doi: 10.6040/j.issn.1671-7554.0.2021.1475
申晓畅1,孙一卿1,颜磊2,赵兴波1
SHEN Xiaochang1, SUN Yiqing1, YAN Lei2, ZHAO Xingbo1
摘要: 目的 探讨芳基烃受体核转位因子样蛋白2(ARNTL2)在子宫内膜癌中的表达、作用及其可能的信号通路。 方法 采用免疫组织化学SP法、RT-PCR和Western blotting实验检测子宫内膜癌及正常内膜组织中ARNTL2的表达;小干扰RNA(Si-ARNTL2)敲低ARNTL2 的表达,CCK-8实验检测细胞增殖能力,划痕愈伤实验和小室穿透实验(Transwell)检测细胞迁移和侵袭能力;Western blotting检测细胞上皮间质转化指标:N-钙黏蛋白(N-cad)、E-钙黏蛋白(E-cad)、Vimentin蛋白、Snail蛋白、Slug蛋白及Wnt/β-catenin信号通路相关蛋白(Wnt蛋白、β-连环蛋白)的表达。 结果 免疫组织化学、RT-PCR和Western blotting结果均显示,ARNTL2在子宫内膜癌组织中表达上调(t免疫组织化学=2.213, P=0.033;tRT-PCR=4.766, P<0.001;tWestern blotting=2.659,P=0.012);Si-ARNTL2转染可成功降低子宫内膜癌细胞系(KLE和Ishikawa)ARNTL2的表达。敲低ARNTL2后,CCK8实验结果显示,两种细胞的吸光度值下降(P均<0.05);划痕愈伤实验结果显示,两种细胞的划痕愈合率下降(P均<0.001);Transwell侵袭及迁移实验结果均显示,两种细胞穿膜细胞数减少(P均<0.01);Western blotting实验结果显示,两细胞系E-cad表达上升(P均<0.05),N-cad、Vimentin、Snail、Slug蛋白表达下降(P均<0.05),两细胞Wnt、β-catenin蛋白的表达均下降(P均<0.05)。 结论 ARNTL2在子宫内膜癌组织中的表达升高,降低子宫内膜癌细胞中ARNTL2的表达可抑制子宫内膜癌的增殖和转移,其可能通过Wnt/β-catenin通路发挥作用。
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[1] Morice P, Leary A, Creutzberg C, et al. Endometrial cancer [J]. Lancet, 2016, 387(10023): 1094-1108. [2] Siegel RL, Miller KD, Fuchs HE. Cancer statistics, 2021 [J]. CA Cancer J Clin, 2021, 71(1): 7-33. [3] Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020 [J]. CA Cancer J Clin, 2020, 70(1): 7-30. [4] Wu H, Dunnett S, Ho YS, et al. The role of sleep deprivation and circadian rhythm disruption as risk factors of Alzheimers disease [J]. Front Neuroendocrinol, 2019, 54: 100764. doi:10.1016/j.yfrne.2019.100764. [5] Zhou J, Wang J, Zhang X, et al. New insights into cancer chronotherapies [J]. Front Pharmacol, 2021, 12: 741295. doi:10.3389/fphar.2021.741295. [6] 中国抗癌协会妇科肿瘤专业委员会.子宫内膜癌诊断与治疗指南(2021年版)[J]. 中国癌症杂志, 2021, 31(6): 501-512. [7] Lu KH, Broaddus RR. Endometrial cancer [J]. N Engl J Med, 2020, 383(21): 2053-2064. [8] Gery S, Koeffler HP. Circadian rhythms and cancer [J]. Cell Cycle, 2010, 9(6): 1097-1103. [9] Crespo M, Leiva M, Sabio G. Circadian clock and liver cancer [J]. Cancers(Basel), 2021, 13(14): 3631. doi: 10.3390/cancers13143631. [10] Yeh KT, Yang MY, Liu TC, et al. Abnormal expression of period 1(PER1)in endometrial carcinoma [J]. J Pathol, 2005, 206(1): 111-120. [11] Wang Z, Wang H, Wang Z, et al. Associated analysis of PER1/TUBB2B with endometrial cancer development caused by circadian rhythm disorders [J]. Med Oncol, 2020, 37(10): 90. doi:10.1007/s12032-020-01415-4. [12] Shih MC, Yeh KT, Tang KP, et al. Promoter methylation in circadian genes of endometrial cancers detected by methylation-specific PCR [J]. Mol Carcinog, 2006, 45(10): 732-740. [13] Takeda N, Maemura K, Horie S, et al. Thrombomodulin is a clock-controlled gene in vascular endothelial cells [J]. J Biol Chem, 2007, 282(45): 32561-32567. [14] Wang Y, Peng S, Liu T, et al. The potential role of clock genes in children attention-deficit/hyperactivity disorder [J]. Sleep Med, 2020, 71: 18-27. doi: 10.1016/j.sleep.2020.02.021. [15] Barbosa Vieira TK, Jurema da Rocha Leão M, Pereira LX, et al. Correlation between circadian rhythm related genes, type 2 diabetes, and cancer: insights from metanalysis of transcriptomics data [J]. Mol Cell Endocrinol, 2021, 526: 111214. doi:10.1016/j.mce.2021.111214. [16] Shi SQ, Ansari TS, McGuinness OP, et al. Circadian disruption leads to insulin resistance and obesity [J]. Curr Biol, 2013, 23(5): 372-381. [17] Azevedo PG, Miranda LR, Nicolau ES, et al. Genetic association of the PERIOD3(PER3)clock gene with extreme obesity [J]. Obes Res Clin Pract, 2021, 15(4): 334-338. [18] Lu M, Huang L, Tang Y, et al. ARNTL2 knockdown suppressed the invasion and migration of colon carcinoma: decreased SMOC2-EMT expression through inactivation of PI3K/AKT pathway [J]. Am J Transl Res, 2020, 12(4): 1293-1308. [19] Brady JJ, Chuang CH, Greenside PG, et al. An arntl2-driven secretome enables lung adenocarcinoma metastatic self-sufficiency [J]. Cancer Cell, 2016, 29(5): 697-710. [20] Wang Z, Liu T, Xue W, et al. ARNTL2 promotes pancreatic ductal adenocarcinoma progression through TGF/BETA pathway and is regulated by miR-26a-5p [J]. Cell Death Dis, 2020, 11(8): 692. doi:10.1038/s41419-020-02839-6. [21] Ha NH, Long J, Cai Q, et al. The circadian rhythm gene Arntl2 is a metastasis susceptibility gene for estrogen receptor-negative breast cancer [J]. PLoS Genet, 2016, 12(9): e1006267. doi:10.1371/journal.pgen.1006267. [22] Pan G, Liu Y, Shang L, et al. EMT-associated microRNAs and their roles in cancer stemness and drug resistance [J]. Cancer Commun(Lond), 2021, 41(3): 199-217. [23] Usman S, Waseem NH, Nguyen TKN, et al. Vimentin is at the heart of epithelial mesenchymal transition(EMT)mediated metastasis [J]. Cancers(Basel), 2021, 13(19): 4985. doi: 10.3390/cancers13194985. [24] Lei Y, Chen L, Zhang G, et al. MicroRNAs target the Wnt/β-catenin signalling pathway to regulate epithelial-mesenchymal transition in cancer(Review)[J]. Oncol Rep, 2020, 44(4): 1299-1313. [25] Rao TP, Kühl M. An updated overview on wnt signalling pathways: a prelude for more [J]. Circ Res, 2010, 106(12): 1798-1806. |
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