[1]冯馨园),张 婷),崔 戈),等.CNRIP1启动子区去甲基化对结肠癌细胞糖酵解及增殖、迁移活性的影响[J].郑州大学学报(医学版),2018,(01):71-75.[doi:10.13705/j.issn.1671-6825.2017.04.074]
 FENG Xinyuan),ZHANG Ting),CUI Ge),et al.Effects of CNRIP1 promoter demethylation on glycolysis, proliferation and migration activity of colon cancer cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2018,(01):71-75.[doi:10.13705/j.issn.1671-6825.2017.04.074]
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CNRIP1启动子区去甲基化对结肠癌细胞糖酵解及增殖、迁移活性的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2018年01期
页码:
71-75
栏目:
应用研究
出版日期:
2018-01-20

文章信息/Info

Title:
Effects of CNRIP1 promoter demethylation on glycolysis, proliferation and migration activity of colon cancer cells
作者:
冯馨园1) 张 婷1) 崔 戈2) 王旗春1) 钱 乙1) 蔡 莹1) 仰舒静1) 张梦荧1) 姚嘉诚1)
1)湖州师范学院医学院病理教研室 浙江湖州 313000 2)湖州师范学院附属第一医院病理科 浙江湖州 313000
Author(s):
FENG Xinyuan1) ZHANG Ting1) CUI Ge2) WANG Qichun1) QIAN Yi1) CAI Ying1) YANG Shujing1) ZHANG Mengying1) YAO Jiacheng1)
1)Department of Pathology, School of Medicine, Huzhou University, Huzhou,Zhejiang 313000 2)Department of Pathology, the First Affiliated Hospital,Huzhou University, Huzhou, Zhejiang 313000
关键词:
结肠癌 大麻素受体互作蛋白1 去甲基化 糖酵解 增殖 迁移
Keywords:
colon cancer cannabinoid receptor interacting protein 1 demethylation glycolysis proliferation migration
分类号:
R735.3
DOI:
10.13705/j.issn.1671-6825.2017.04.074
摘要:
目的:探讨CNRIP1启动子区去甲基化对结肠癌细胞糖酵解途径、细胞增殖及迁移活性的影响。方法:采用5-氮杂胞苷(5-azaC)降低结肠癌细胞SW620 CNRIP1启动子区甲基化水平,以未干预细胞作对照。用亚硫酸盐测序法检测细胞CNRIP1启动子区甲基化水平; 分别采用qRT-PCR、Western blot法检测细胞中CNRIP1、糖酵解途径关键酶HK2、PKM2、LDH-A mRNA及蛋白表达水平,生化法检测细胞培养液中乳酸水平; 分别采用CCK-8比色法、Transwell法检测细胞增殖及迁移能力。结果:与未干预组细胞相比,5-azaC干预组细胞CNRIP1启动子区甲基化水平降低,CNRIP1 mRNA及蛋白表达升高,HK2、PKM2、LDH-A mRNA和蛋白表达水平及细胞培养液中乳酸水平均降低,细胞增殖及迁移能力均减弱(P均<0.05)。结论:CNRIP1去甲基化可抑制糖酵解途径,从而降低结肠癌细胞的增殖及迁移能力。
Abstract:
Aim: To investigate the effects of cannabinoid receptor interacting protein 1(CNRIP1)promoter demethylation on glycolysis, proliferation, and migration potential of colon cancer cells.Methods: CNRIP1 promoter methylation in colon cancer cells SW620 was suppressed via treatment with 5-azacytidine(5-azaC), and the CNRIP1 promoter methylation level was determined by bisulfite sequencing PCR; the mRNA and protein expression levels of CNRIP1 as well as key enzymes(HK2, PKM2, LDH-A)of glycolysis pathway were determined by qRT-PCR and Western blot, respectively; lactic acid level in culture medium was determined with biochemistry kit. The proliferative and migration abilities of colon cancer cells were determined with CCK-8 and Transwell cell assays, respectively.Results: Compared with untreated group, CNRIP1 promoter methylation level was decreased significantly in SW620 cells treated with 5-azaC, mRNA and protein expressions of CNRIP1 were increased significantly, mRNA and protein expressions of HK2, PKM2 and LDH-A were depressed significantly, and the proliferative and migration abilities were depressed significantly(P<0.05).Conclusion: CNRIP1 promoter demethylation could depress the glycolysis pathway, and reduce the proliferative and migration abilities in colon cancer cells.

参考文献/References:

[1] SMITH TH,BLUME LC,STRAIKER A,et al.Cannabinoid receptor-interacting protein 1a modulates CB1 receptor signaling and regulation[J].Mol Pharmacol,2015,87(4):747
[2] ZHENG X,SUZUKI T,TAKAHASHI C,et al.cnrip1 is a regulator of eye and neural development in Xenopus laevis[J].Genes Cells,2015,20(4):324
[3] SMITH TH,SIM-SELLEY LJ,SELLEY DE.Cannabinoid CB1 receptor-interacting proteins: novel targets for central nervous system drug discovery?[J].Br J Pharmacol,2010,160(3):454
[4] LIND GE,DANIELSEN SA,AHLQUIST T,et al.Identification of an epigenetic biomarker panel with high sensitivity and specificity for colorectal cancer and adenomas[J].Mol Cancer,2011,10:85
[5] OSTER B,THORSEN K,LAMY P,et al.Identification and validation of highly frequent CpG island hypermethylation in colorectal adenomas and carcinomas[J].Int J Cancer,2011,129(12):2855
[6] BETHGE N,LOTHE RA,HONNE H,et al.Colorectal cancer DNA methylation marker panel validated with high performance in Non-Hodgkin lymphoma[J].Epigenetics,2014,9(3):428
[7] KULIS M,ESTELLER M.DNA methylation and cancer[J].Adv Genet,2010,70:27
[8] CHEISHVILI D,BOUREAU L,SZYF M.DNA demethylation and invasive cancer: implications for therapeutics[J].Br J Pharmacol,2015,172(11):2705
[9] 张婷,崔戈,冯文明,等.结直肠腺癌患者外周血CNRIP1基因启动子甲基化的临床病理学分析[J].临床与实验病理学杂志,2014,30(10):1086
[10]BARRES R,ZIERATH JR.DNA methylation in metabolic disorders[J].Am J Clin Nutr,2011,93(4):897S
[11]WARBURG O.On the origin of cancer cells[J].Science,1956,123(3191):309
[12]PATRA KC,HAY N.Hexokinase 2 as oncotarget[J].Oncotarget,2013,4(11):1862
[13]LI C,ZHAO Z,ZHOU Z,et al.PKM2 promotes cell survival and invasion under metabolic stress by enhancing Warburg effect in pancreatic ductal adenocarcinoma[J].Dig Dis Sci,2016,61(3):767
[14]MIAO P,SHENG S,SUN X,et al.Lactate dehydrogenase A in cancer:a promising target for diagnosis and therapy[J].IUBMB Life,2013,65(11):904

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备注/Memo

备注/Memo:
【基金项目】浙江省自然科学基金项目(LQ16C080001); 浙江省医药卫生研究计划项目(2015KYB372); 湖州市科技计划项目(2015GZ14); 国家级大学生创新训练项目(201610347015)
【作者简介】张婷,通信作者,女,1981年12月生,硕士,副教授,研究方向:肿瘤病理学,E-mail:grapechang@163.com
更新日期/Last Update: 2018-01-20